Solid forms comprising (+)-2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione, compositions thereof, and uses thereof

ABSTRACT

Solid forms comprising (+)-2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione, compositions comprising the solid forms, methods of making the solid forms and methods of their use are disclosed. The methods include methods of treating and/or preventing disorders ameliorated by the reduction of levels of TNF-α or the inhibition of PDE4.

This application is a continuation of U.S. patent application Ser. No.12/079,615, filed Mar. 27, 2008, which is a continuation-in-part of U.S.patent application Ser. No. 11/106,142, filed Apr. 13, 2005, which is adivisional of U.S. patent application Ser. No. 10/392,195, filed on Mar.19, 2003, issued as U.S. Pat. No. 6,962,940, which claims the benefit ofU.S. Provisional Patent Application No. 60/366,515, filed on Mar. 20,2002, and U.S. Provisional Patent Application No. 60/438,450, filed onJan. 7, 2003, the entireties of which are incorporated herein byreference.

1. FIELD OF INVENTION

Provided herein are solid forms comprising(+)-2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione,compositions comprising the solid forms, methods of making the solidforms and methods of their use for the treatment of various diseasesand/or disorders.

2. BACKGROUND OF THE INVENTION

Tumor necrosis factor alpha (TNF-α) is a cytokine that is releasedprimarily by mononuclear phagocytes in response to immunostimulators.TNF-α is capable of enhancing most cellular processes, such asdifferentiation, recruitment, proliferation, and proteolyticdegradation. At low levels, TNF-α confers protection against infectiveagents, tumors, and tissue damage. However, TNF-α also has a role inmany diseases. When administered to a patient, TNF-α causes oraggravates inflammation, fever, cardiovascular effects, hemorrhage,coagulation, and acute phase responses similar to those seen duringacute infections and shock states. Enhanced or unregulated TNF-αproduction has been implicated in a number of diseases and medicalconditions, for example, cancers, such as solid tumors and blood-bornetumors; heart disease, such as congestive heart failure; and viral,genetic, inflammatory, allergic, and autoimmune diseases.

Adenosine 3′,5′-cyclic monophosphate (cAMP) also plays a role in manydiseases and conditions, such as, but not limited to, asthma andinflammation, and other conditions (Lowe and Cheng, Drugs of the Future,17(9), 799-807, 1992). It has been shown that the elevation of cAMP ininflammatory leukocytes inhibits their activation and the subsequentrelease of inflammatory mediators, including TNF-α and NF-κB. Increasedlevels of cAMP also leads to the relaxation of airway smooth muscle.

It is believed that the primary cellular mechanism for the inactivationof cAMP is the breakdown of cAMP by a family of isoenzymes referred toas cyclic nucleotide phosphodiesterases (PDE) (Beavo and Reitsnyder,Trends in Pharm., 11, 150-155, 1990). There are eleven known PDEfamilies. It is recognized, for example, that the inhibition of PDE typeIV is particularly effective in both the inhibition of inflammatorymediator release and the relaxation of airway smooth muscle (Verghese,et al., J. Pharm. Exper. Therapeut., 272(3), 1313-1320, 1995). Thus,compounds that inhibit PDE4 (PDE IV) specifically, may inhibitinflammation and aid the relaxation of airway smooth muscle with aminimum of unwanted side effects, such as cardiovascular oranti-platelet effects. Currently used PDE4 inhibitors lack the selectiveaction at acceptable therapeutic doses.

Cancer is a particularly devastating disease, and increases in bloodTNF-α levels are implicated in the risk of and the spreading of cancer.Normally, in healthy subjects, cancer cells fail to survive in thecirculatory system, one of the reasons being that the lining of bloodvessels acts as a barrier to tumor-cell extravasation. However,increased levels of cytokines have been shown to substantially increasethe adhesion of cancer cells to endothelium in vitro. One explanation isthat cytokines, such as TNF-α, stimulate the biosynthesis and expressionof a cell surface receptors called ELAM-1 (endothelial leukocyteadhesion molecule). ELAM-1 is a member of a family of calcium-dependentcell adhesion receptors, known as LEC-CAMs, which includes LECAM-1 andGMP-140. During an inflammatory response, ELAM-1 on endothelial cellsfunctions as a “homing receptor” for leukocytes. Recently, ELAM-1 onendothelial cells was shown to mediate the increased adhesion of coloncancer cells to endothelium treated with cytokines (Rice et al., 1989,Science 246:1303-1306).

Inflammatory diseases such as arthritis, related arthritic conditions(e.g., osteoarthritis and rheumatoid arthritis), inflammatory boweldisease (e.g., Crohn's disease and ulcerative colitis), sepsis,psoriasis, atopic dermatitis, contact dermatitis, chronic obstructivepulmonary disease, and chronic inflammatory pulmonary diseases are alsoprevalent and problematic ailments. TNF-α plays a central role in theinflammatory response and the administration of their antagonists blockchronic and acute responses in animal models of inflammatory disease.

Enhanced or unregulated TNF-α production has been implicated in viral,genetic, inflammatory, allergic, and autoimmune diseases. Examples ofsuch diseases include but are not limited to: HIV; hepatitis; adultrespiratory distress syndrome; bone-resorption diseases; chronicobstructive pulmonary diseases; chronic pulmonary inflammatory diseases;asthma; dermatitis; cystic fibrosis; septic shock; sepsis; endotoxicshock; hemodynamic shock; sepsis syndrome; post ischemic reperfusioninjury; meningitis; psoriasis; fibrotic disease; cachexia; graftrejection; auto-immune disease; rheumatoid spondylitis; arthriticconditions, such as rheumatoid arthritis and osteoarthritis;osteoporosis; Crohn's disease; ulcerative colitis; inflammatory-boweldisease; multiple sclerosis; systemic lupus erythrematosus; ENL inleprosy; radiation damage; asthma; and hyperoxic alveolar injury. Traceyet al., 1987, Nature 330:662-664 and Hinshaw et al., 1990, Circ. Shock30:279-292 (endotoxic shock); Dezube et al., 1990, Lancet, 335:662(cachexia); Millar et al., 1989, Lancet 2:712-714 and Ferrai-Balivieraet al., 1989, Arch. Surg. 124:1400-1405 (adult respiratory distresssyndrome); Bertolini et al., 1986, Nature 319:516-518, Johnson et al.,1989, Endocrinology 124:1424-1427, Holler et al., 1990, Blood75:1011-1016, and Grau et al., 1989, N. Engl. J. Med. 320:1586-1591(bone resorption diseases); Pignet et al., 1990, Nature, 344:245-247,Bissonnette et al., 1989, Inflammation 13:329-339 and Baughman et al.,1990, J. Lab. Clin. Med. 115:36-42 (chronic pulmonary inflammatorydiseases); Elliot et al., 1995, Int. J. Pharmac. 17:141-145 (rheumatoidarthritis); von Dullemen et al., 1995, Gastroenterology, 109:129-135(Crohn's disease); Duh et al., 1989, Proc. Nat. Acad. Sci. 86:5974-5978,Poll et al., 1990, Proc. Nat. Acad. Sci. 87:782-785, Monto et al., 1990,Blood 79:2670, Clouse et al., 1989, J. Immunol. 142, 431-438, Poll etal., 1992, AIDS Res. Hum. Retrovirus, 191-197, Poli et al. 1990, Proc.Natl. Acad. Sci. 87:782-784, Folks et al., 1989, PNAS 86:2365-2368 (HIVand opportunistic infections resulting from HIV).

Pharmaceutical compounds that can block the activity or inhibit theproduction of certain cytokines, including TNF-α, may be beneficialtherapeutics. Many small-molecule inhibitors have demonstrated anability to treat or prevent inflammatory diseases implicated by TNF-α(for a review, see Lowe, 1998 Exp. Opin. Ther. Patents 8:1309-1332). Onesuch class of molecules are the substituted phenethylsulfones describedin U.S. Pat. No. 6,020,358.

The preparation and selection of a solid form of a pharmaceuticalcompound is complex, given that a change in solid form may affect avariety of physical and chemical properties, which may provide benefitsor drawbacks in processing, formulation, stability and bioavailability,among other important pharmaceutical characteristics. Potentialpharmaceutical solids include crystalline solids and amorphous solids.Amorphous solids are characterized by a lack of long-range structuralorder, whereas crystalline solids are characterized by structuralperiodicity. The desired class of pharmaceutical solid depends upon thespecific application; amorphous solids are sometimes selected on thebasis of e.g., an enhanced dissolution profile, while crystalline solidsmay be desirable for properties such as, e.g., physical or chemicalstability (see, e.g., S. R. Vippagunta et al., Adv. Drug. Deliv. Rev.,(2001) 48:3-26; L. Yu, Adv. Drug. Deliv. Rev., (2001) 48:27-42).

Whether crystalline or amorphous, potential solid forms of apharmaceutical compound include single-component and multiple-componentsolids. Single-component solids consist essentially of thepharmaceutical compound in the absence of other compounds. Variety amongsingle-component crystalline materials may potentially arise, e.g., fromthe phenomenon of polymorphism, wherein multiple three-dimensionalarrangements exist for a particular pharmaceutical compound (see, e.g.,S. R. Byrn et al., Solid State Chemistry of Drugs, (1999) SSCI, WestLafayette). The importance of studying polymorphs was underscored by thecase of Ritonavir, an HIV protease inhibitor that was formulated as softgelatin capsules. About two years after the product was launched, theunanticipated precipitation of a new, less soluble polymorph in theformulation necessitated the withdrawal of the product from the marketuntil a more consistent formulation could be developed (see S. R.Chemburkar et al., Org. Process Res. Dev., (2000) 4:413-417).

Additional diversity among the potential solid forms of a pharmaceuticalcompound may arise, e.g., from the possibility of multiple-componentsolids. Crystalline solids comprising two or more ionic species may betermed salts (see, e.g., Handbook of Pharmaceutical Salts Properties,Selection and Use, P. H. Stahl and C. G. Wermuth, Eds., (2002), Wiley,Weinheim). Additional types of multiple-component solids that maypotentially offer other property improvements for a pharmaceuticalcompound or salt thereof include, e.g., hydrates, solvates, co-crystalsand clathrates, among others (see, e.g., S. R. Byrn et al., Solid StateChemistry of Drugs, (1999) SSCI, West Lafayette). Moreover,multiple-component crystal forms may potentially be susceptible topolymorphism, wherein a given multiple-component composition may existin more than one three-dimensional crystalline arrangement. Thepreparation of solid forms is of great importance in the development ofa safe, effective, stable and marketable pharmaceutical compound.

Provided herein are embodiments addressing a need for solid forms of thecompound chemically named(+)-2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione(“Compound A”), which was disclosed in U.S. application Ser. No.10/392,195, filed Mar. 19, 2003 (issued as U.S. Pat. No. 6,962,940), aswell as U.S. Provisional Application Ser. Nos. 60/366,515, filed Mar.20, 2002 and 60/438,450, filed Jan. 7, 2003.

3. SUMMARY OF THE INVENTION

This invention relates to methods of treating diseases and disordersutilizing an enantiomer of a substituted phenethylsulfone compound andpharmaceutically acceptable solvates, hydrates, co-crystals, clathrates,prodrugs and polymorphs thereof and methods for reducing the level ofcytokines and their precursors in mammals. The invention also relates topharmaceutical compositions comprising the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneand a pharmaceutically acceptable carrier. The invention further relatesto the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dionesubstantially free of its (−) enantiomer.

This invention particularly relates to the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.This compound is believed to have increased potency and other benefitsas compared to its racemate,2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.

The invention encompasses the use of the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dionefor treating or preventing diseases or disorders ameliorated by theinhibition of TNF-α production in mammals. In certain embodiments, thistreatment includes the reduction or avoidance of adverse effects. Suchdisorders include, but are not limited to, cancers, including, but notlimited to cancer of the head, thyroid, neck, eye, skin, mouth, throat,esophagus, chest, bone, blood, bone marrow, lung, colon, sigmoid,rectum, stomach, prostate, breast, ovaries, kidney, liver, pancreas,brain, intestine, heart, adrenal, subcutaneous tissue, lymph nodes,heart, and combinations thereof. Specific cancers that can be treated bythis method are multiple myeloma, malignant melanoma, malignant glioma,leukemia and solid tumors.

The invention also encompasses the use of the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dionein the treatment or prevention of heart disease, including, but notlimited to congestive heart failure, cardiomyopathy, pulmonary edema,endotoxin-mediated septic shock, acute viral myocarditis, cardiacallograft rejection, and myocardial infarction.

The invention also encompasses the use of the (+) enantiomer of2-[1-[3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneto treat diseases or disorders ameliorated by the inhibition of PDE4.For example, the compounds and compositions of the invention may beuseful to treat or prevent viral, genetic, inflammatory, allergic, andautoimmune diseases. Examples of such diseases include, but are notlimited to: HIV; hepatitis; adult respiratory distress syndrome;bone-resorption diseases; chronic obstructive pulmonary diseases;chronic pulmonary inflammatory diseases; dermatitis; inflammatory skindisease, atopic dermatitis, cystic fibrosis; septic shock; sepsis;endotoxic shock; hemodynamic shock; sepsis syndrome; post ischemicreperfusion injury; meningitis; psoriasis; fibrotic disease; cachexia;graft rejection including graft versus host disease; auto-immunedisease; rheumatoid spondylitis; arthritic conditions, such asrheumatoid arthritis and osteoarthritis; osteoporosis; Crohn's disease;ulcerative colitis; inflammatory-bowel disease; multiple sclerosis;systemic lupus erythrematosus; erythema nodosum leprosum (ENL) inleprosy; radiation damage; asthma; and hyperoxic alveolar injury.

In yet another embodiment, the stereomerically pure (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneis also useful in the treatment or prevention of microbial infections orthe symptoms of microbial infections including, but not limited to,bacterial infections, fungal infections, malaria, mycobacterialinfection, and opportunistic infections resulting from HTV.

The invention further encompasses pharmaceutical compositions and singleunit dosage forms comprising the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneand pharmaceutically acceptable polymorphs, prodrugs, hydrates,clathrates, and solvates thereof.

In a separate embodiment, the invention encompasses the (+) enantiomerof2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.

In a further embodiment, the invention encompasses a method of producingthe stereomerically pure (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dionewhich comprises contacting1-(3-Ethoxy-4-methoxy-phenyl)-2-methanesulfonyl-ethylamine with a chiralamino acid and contacting the product of the first step withN-(1,3-Dioxo-1,3-dihydro-isobenzofuran-4-yl)-acetamide. In a relatedembodiment the invention encompasses a chiral salt of1-(3-Ethoxy-4-methoxy-phenyl)-2-methanesulfonyl-ethylamine.

Embodiments herein provide solid forms comprising the compoundchemically named(+)-2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione(“Compound A”). Compound A can be synthesized or obtained according toany method apparent to those of skill in the art based upon theteachings herein, including the methods described in the Examples below.Compound A can also be prepared according to the methods described inU.S. Pat. No. 6,962,940, issued Nov. 8, 2005, the entirety of which isincorporated by reference herein.

In certain embodiments, the solid forms are single-component crystalforms of Compound A. In certain embodiments, the solid forms aremultiple-component crystal forms, including, but not limited to,co-crystals and/or solvates (including hydrates) comprising Compound A.In other embodiments, the solid forms are single-component amorphousforms of Compound A. In other embodiments, the solid forms aremultiple-component amorphous forms. Without intending to be limited byany particular theory, certain novel solid forms provided herein haveparticular advantageous physical and/or chemical properties making themuseful, e.g., for manufacturing, processing, formulation and/or storage,while also possessing particularly advantageous biological properties,such as, e.g., bioavailability and/or biological activity.

In particular embodiments, solid forms provided herein include solidforms comprising Compound A, including, but not limited to,single-component and multiple-component solid forms comprising CompoundA. In certain embodiments, solid forms provided herein includepolymorphs, solvates (including hydrates) and co-crystals comprisingCompound A. Certain embodiments herein provide methods of making,isolating and/or characterizing the solid forms provided herein.

The solid forms provided herein are useful as active pharmaceuticalingredients for the preparation of formulations for use in patients.Thus, embodiments herein encompass the use of these solid forms as afinal drug product. Certain embodiments provide solid forms useful inmaking final dosage forms with improved properties, e.g., powder flowproperties, compaction properties, tableting properties, stabilityproperties, and excipient compatibility properties, among others, thatare needed for manufacturing, processing, formulation and/or storage offinal drug products. Certain embodiments herein provide pharmaceuticalcompositions comprising a single-component crystal form, amultiple-component crystal form, a single-component amorphous formand/or a multiple-component amorphous form comprising Compound A and apharmaceutically acceptable diluent, excipient or carrier. The solidforms and the final drug products provided herein are useful, forexample, for the treatment, prevention or management of diseases anddisorders provided herein.

Certain embodiments herein provide methods using the solid formsprovided herein for treating, preventing or managing diseases ordisorders ameliorated by the inhibition of TNF-α production in mammals,such as HIV; hepatitis; adult respiratory distress syndrome; boneresorption diseases; chronic obstructive pulmonary diseases; chronicpulmonary inflammatory diseases; asthma; dermatitis; cystic fibrosis;septic shock; sepsis; endotoxic shock; hemodynamic shock; sepsissyndrome; post ischemic reperfusion injury; meningitis; psoriasis;fibrotic disease; cachexia; graft rejection; auto immune disease;rheumatoid spondylitis; arthritic conditions, such as psoriaticarthritis, rheumatoid arthritis and osteoarthritis; osteoporosis;Crohn's disease; ulcerative colitis; inflammatory bowel disease;multiple sclerosis; systemic lupus erythematosus; cutaneous lupuserythematosus; pulmonary sarcoidosis; ENL in leprosy; radiation damage;asthma; and hyperoxic alveolar injury. Such disorders further include,but are not limited to, cancers, including, but not limited to cancer ofthe head, thyroid, neck, eye, skin, mouth, throat, esophagus, chest,bone, blood, bone marrow, lung, colon, sigmoid, rectum, stomach,prostate, breast, ovaries, kidney, liver, pancreas, brain, intestine,heart, adrenal, subcutaneous tissue, lymph nodes, heart, andcombinations thereof. Specific cancers that can be treated by thismethod are multiple myeloma, malignant melanoma, malignant glioma,leukemia and solid tumors. In certain embodiments, methods using thesolid forms provided herein include the reduction or avoidance ofcertain adverse effects.

Certain embodiments herein provide methods of using the solid formsprovided herein in the treatment or prevention of heart disease,including, but not limited to congestive heart failure, cardiomyopathy,pulmonary edema, endotoxin-mediated septic shock, acute viralmyocarditis, cardiac allograft rejection, and myocardial infarction.

Certain embodiments herein provide methods of using the solid formsprovided herein to treat diseases or disorders ameliorated by theinhibition of PDE4. For example, the solid forms provided herein may beuseful to treat or prevent viral, genetic, inflammatory, allergic, andautoimmune diseases. Examples of such diseases include, but are notlimited to: HIV; hepatitis; adult respiratory distress syndrome;bone-resorption diseases; chronic obstructive pulmonary diseases;chronic pulmonary inflammatory diseases; dermatitis; inflammatory skindisease; atopic dermatitis; cystic fibrosis; septic shock; sepsis;endotoxic shock; hemodynamic shock; sepsis syndrome; post ischemicreperfusion injury; meningitis; psoriasis; fibrotic disease; cachexia;graft rejection including graft versus host disease; auto-immunedisease; rheumatoid spondylitis; arthritic conditions, such asrheumatoid arthritis and osteoarthritis; osteoporosis; Crohn's disease;ulcerative colitis; inflammatory-bowel disease; multiple sclerosis;systemic lupus erythrematosus; erythema nodosum leprosum (ENL) inleprosy; radiation damage; asthma; and hyperoxic alveolar injury.

Certain embodiments herein provide methods of using the solid formsprovided herein in the treatment or prevention of microbial infectionsor the symptoms of microbial infections including, but not limited to,bacterial infections, fungal infections, malaria, mycobacterialinfection, and opportunistic infections resulting from HIV.

Particular embodiments herein provide methods of using the solid formsprovided herein in the treatment or prevention of diseases including:psoriasis; psoriatic arthritis; rheumatoid arthritis; chronic cutaneoussarcoid; giant cell arteritis; Parkinson's; prurigo nodularis; lichenplanus; complex apthosis; Behcet's disease; lupus; hepatitis; uveitis;Sjogren's disease; depression (including major depression); interstitialcystitis; vulvodynia; prostatitis; osteoarthritis; diffuse large B celllymphoma; polymysoitis; dermatomyositis; inclusiuon body myositis;erosive osteoarthritis; interstitial cystitis; hepatitis; endometriosis;radiculopathy; and pyoderma gangrenosum.

Certain embodiments herein provide pharmaceutical compositions andsingle unit dosage forms comprising one or more solid forms providedherein.

3.1. BRIEF DESCRIPTION OF THE FIGURES

FIG. 1 provides a representative X-ray Powder Diffraction (“XRPD”)pattern of Form A of Compound A.

FIG. 2 provides a representative Differential Scanning calorimetry(“DSC”) plot of Form A of Compound A.

FIG. 3 provides a representative Thermal Gravimetric Analysis (“TGA”)plot of Form A of Compound A.

FIG. 4 provides a representative Dynamic Vapor Sorption (“DVS”) plot ofForm A of Compound A.

FIG. 5 provides a representative XRPD pattern of Form B of Compound A.

FIG. 6 provides a representative DSC plot of Form B of Compound A.

FIG. 7 provides a representative TGA plot of Form B of Compound A.

FIG. 8 provides a representative DVS plot of Form B of Compound A.

FIG. 9 provides a representative XRPD pattern of Form C of Compound A.

FIG. 10 provides a representative DSC plot of Form C of Compound A.

FIG. 11 provides a representative TGA plot of Form C of Compound A.

FIG. 12 provides a representative DVS plot of Form C of Compound A.

FIG. 13 provides a representative XRPD pattern of Form D of Compound A.

FIG. 14 provides a representative DSC plot of Form D of Compound A.

FIG. 15 provides a representative TGA plot of Form D of Compound A.

FIG. 16 provides a representative DVS plot of Form D of Compound A.

FIG. 17 provides a representative XRPD pattern of Form E of Compound A.

FIG. 18 provides a representative DSC plot of Form E of Compound A.

FIG. 19 provides a representative TGA plot of Form E of Compound A.

FIG. 20 provides a representative DVS plot of Form E of Compound A.

FIG. 21 provides a representative XRPD pattern of Form F of Compound A.

FIG. 22 provides a representative DSC plot of Form F of Compound A.

FIG. 23 provides a representative TGA plot of Form F of Compound A.

FIG. 24 provides a representative DVS plot of Form F of Compound A.

FIG. 25 provides a representative XRPD of Form G of Compound A.

FIG. 26 provides a representative DSC plot of Form G of Compound A.

FIG. 27 provides a representative TGA plot of Form G of Compound A.

FIG. 28 provides a representative DVS plot of Form G of Compound A.

FIG. 29 illustrates a preparation of the (+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.

FIG. 30 illustrates the effect of Compound A on LPS-induced neutrophiliain the lungs of conscious ferrets.

FIG. 31 illustrates the percent change in epidermal thickness among all15 subjects at Day 29 in a clinical study evaluating Compound A inpatients with severe plaque-type psoriasis.

FIG. 32 illustrates the change in mean iNOS (normalized to hARP) inbiopsy specimens of lesional skin at Day 29 in a clinical studyevaluating Compound A in patients with severe plaque-type psoriasis.

FIG. 33 illustrates the percentage change in total Psoriasis Area andSeverity Index (PASO score among evaluable patients from baseline at Day29 in a clinical study evaluating Compound A in patients with severeplaque-type psoriasis.

3.2. DEFINITIONS

As used herein, term “Compound A” refers to enantiomerically pure(+)-2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dionewhich comes off of an HPLC column at about 25.4 minutes when that columnis a 150 mm×4.6 mm Ultron Chiral ES-OVS chiral HPLC column (AgilentTechnology), the eluent is 15:85 ethanol: 20 mM KH₂PO₄ at pH 3.5, andthe observation wavelength is 240 nm. The ¹H NMR spectrum of Compound Ais substantially as follows: δ(CDCl₃); 1.47 (t, 3H); 2.26 (s, 3H); 2.87(s, 3H); 3.68-3.75 (dd, 1H); 3.85 (s, 3H); 4.07-4.15 (q, 2H); 4.51-4.61(dd, 1H); 5.84-5.90 (dd, 1H); 6.82-8.77 (m, 6H); 9.46 (s, 1H). The ¹³CNMR spectrum of Compound A is substantially as follows: δ(DMSO-d₆);14.66; 24.92; 41.61; 48.53; 54.46; 55.91; 64.51; 111.44; 112.40; 115.10;118.20; 120.28; 124.94; 129.22; 131.02; 136.09; 137.60; 148.62; 149.74;167.46; 169.14; 169.48. Compound A dissolved in methanol rotates planepolarized light in the (+) direction.

Without being limited by theory, Compound A is believed to beS-{2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione},which has the following structure:

As used herein, the term “patient” refers to a mammal, particularly ahuman.

As used herein, the term “pharmaceutically acceptable salts” refer tosalts prepared from pharmaceutically acceptable non-toxic acids or basesincluding inorganic acids and bases and organic acids and bases.

As used herein and unless otherwise indicated, the term “prodrug” meansa derivative of a compound that can hydrolyze, oxidize, or otherwisereact under biological conditions (in vitro or in vivo) to provide thecompound. Examples of prodrugs include, but are not limited to,derivatives and metabolites of Compound A that include biohydrolyzablemoieties such as biohydrolyzable amides, biohydrolyzable esters,biohydrolyzable carbamates, biohydrolyzable carbonates, biohydrolyzableureides, and biohydrolyzable phosphate analogues. Prodrugs can typicallybe prepared using well-known methods, such as those described by 1Burger's Medicinal Chemistry and Drug Discovery, 172-178, 949-982(Manfred E. Wolff ed., 5th ed. 1995).

As used herein and unless otherwise indicated, the terms“biohydrolyzable amide,” “biohydrolyzable ester,” “biohydrolyzablecarbamate,” “biohydrolyzable carbonate,” “biohydrolyzable ureide,”“biohydrolyzable phosphate” mean an amide, ester, carbamate, carbonate,ureide, or phosphate, respectively, of a compound that either: 1) doesnot interfere with the biological activity of the compound but canconfer upon that compound advantageous properties in vivo, such asuptake, duration of action, or onset of action; or 2) is biologicallyinactive but is converted in vivo to the biologically active compound.Examples of biohydrolyzable esters include, but are not limited to,lower alkyl esters, alkoxyacyloxy esters, alkyl acylamino alkyl esters,and choline esters. Examples of biohydrolyzable amides include, but arenot limited to, lower alkyl amides, α-amino acid amides, alkoxyacylamides, and alkylaminoalkylcarbonyl amides. Examples of biohydrolyzablecarbamates include, but are not limited to, lower alkylamines,substituted ethylenediamines, aminoacids, hydroxyalkylamines,heterocyclic and heteroaromatic amines, and polyether amines.

As used herein and unless otherwise indicated, the term “stereomericallypure” means a composition that comprises one stereoisomer of a compoundand is substantially free of other stereoisomers of that compound. Forexample, a stereomerically pure composition of a compound having onechiral center will be substantially free of the opposite enantiomer ofthe compound. A stereomerically pure composition of a compound havingtwo chiral centers will be substantially free of other diastereomers ofthe compound. A typical stereomerically pure compound comprises greaterthan about 80% by weight of one stereoisomer of the compound and lessthan about 20% by weight of other stereoisomers of the compound, morepreferably greater than about 90% by weight of one stereoisomer of thecompound and less than about 10% by weight of the other stereoisomers ofthe compound, even more preferably greater than about 95% by weight ofone stereoisomer of the compound and less than about 5% by weight of theother stereoisomers of the compound, and most preferably greater thanabout 97% by weight of one stercoisomer of the compound and less thanabout 3% by weight of the other stereoisomers of the compound.

As used herein and unless otherwise indicated, the term“enantiomerically pure” means a stereomerically pure composition of acompound having one chiral center.

As used herein, term “adverse effects” includes, but is not limited togastrointestinal, renal and hepatic toxicities, leukopenia, increases inbleeding times due to, e.g., thrombocytopenia, and prolongation ofgestation, nausea, vomiting, somnolence, asthenia, dizziness,teratogenicity, extra-pyramidal symptoms, akathisia, cardiotoxicityincluding cardiovascular disturbances, inflammation, male sexualdysfunction, and elevated scrum liver enzyme levels. The term“gastrointestinal toxicities” includes but is not limited to gastric andintestinal ulcerations and erosions. The term “renal toxicities”includes but is not limited to such conditions as papillary necrosis andchronic interstitial nephritis.

As used herein and unless otherwise indicated, the phrases “reduce oravoid adverse effects” and “reducing or avoiding adverse effects” meanthe reduction of the severity of one or more adverse effects as definedherein.

It should be noted that if there is a discrepancy between a depictedstructure and a name given that structure, the depicted structure is tobe accorded more weight. In addition, if the stereochemistry of astructure or a portion of a structure is not indicated with, forexample, bold or dashed lines, the structure or portion of the structureis to be interpreted as encompassing all stereoisomers of it.

As used herein and unless otherwise specified, the terms “solid form”and related terms refer to a physical form which is not predominantly ina liquid or a gaseous state. As used herein and unless otherwisespecified, the term “solid form” and related terms, when used herein torefer to Compound A, refer to a physical form comprising Compound Awhich is not predominantly in a liquid or a gaseous state. Solid formsmay be crystalline, amorphous or mixtures thereof. In particularembodiments, solid forms may be liquid crystals. A “single-component”solid form comprising Compound A consists essentially of Compound A. A“multiple-component” solid form comprising Compound A comprises asignificant quantity of one or more additional species, such as ionsand/or molecules, within the solid form. For example, in particularembodiments, a crystalline multiple-component solid form comprisingCompound A further comprises one or more species non-covalently bondedat regular positions in the crystal lattice. Multiple-component solidforms comprising Compound A include co-crystals, solvates (e.g.,hydrates), and clathrates of Compound A. In particular embodiments, theterm “solid form comprising Compound A” and related terms includesingle-component and multiple-component solid forms comprising CompoundA. In particular embodiments, “solid forms comprising Compound A” andrelated terms include crystal forms comprising Compound A, amorphousforms comprising Compound A, and mixtures thereof.

As used herein and unless otherwise specified, the term “crystalline”and related terms used herein, when used to describe a compound,substance, modification, material, component or product, unlessotherwise specified, mean that the compound, substance, modification,material, component or product is substantially crystalline asdetermined by X-ray diffraction. See, e.g., Remington: The Science andPractice of Pharmacy, 21^(st) edition, Lippincott, Williams and Wilkins,Baltimore, Md. (2005); The United States Pharmacopeia, 23^(rd) ed.,1843-1844 (1995).

As used herein and unless otherwise specified, the term “crystal forms,”“crystalline forms” and related terms herein refer to solid forms thatare crystalline. Crystal forms include single-component crystal formsand multiple-component crystal forms, and include, but are not limitedto, polymorphs, solvates, hydrates, and/or other molecular complexes. Incertain embodiments, a crystal form of a substance may be substantiallyfree of amorphous forms and/or other crystal forms. In certainembodiments, a crystal form of a substance may contain less than about1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 15%, 20%, 25%, 30%, 35%, 40%,45% or 50% of one or more amorphous forms and/or other crystal forms ona weight basis. In certain embodiments, a crystal form of a substancemay be physically and/or chemically pure. In certain embodiments, acrystal form of a substance may be about 99%, 98%, 97%, 96%, 95%, 94%,93%, 92%, 91% or 90% physically and/or chemically pure.

As used herein and unless otherwise specified, the terms “polymorphs,”“polymorphic forms” and related terms herein, refer to two or morecrystal forms that consist essentially of the same molecule, molecules,and/or ions. Like different crystal forms, different polymorphs may havedifferent physical properties such as, e.g., melting temperature, heatof fusion, solubility, dissolution properties and/or vibrationalspectra, as a result of the arrangement or conformation of the moleculesand/or ions in the crystal lattice. The differences in physicalproperties may affect pharmaceutical parameters such as storagestability, compressibility and density (important in formulation andproduct manufacturing), and dissolution rate (an important factor inbioavailability). Differences in stability can result from changes inchemical reactivity (e.g., differential oxidation, such that a dosageform discolors more rapidly when comprised of one polymorph than whencomprised of another polymorph) or mechanical changes (e.g., tabletscrumble on storage as a kinetically favored polymorph converts tothermodynamically more stable polymorph) or both (e.g., tablets of onepolymorph are more susceptible to breakdown at high humidity). As aresult of solubility/dissolution differences, in the extreme case, somesolid-state transitions may result in lack of potency or, at the otherextreme, toxicity. In addition, the physical properties may be importantin processing (e.g., one polymorph might be more likely to form solvatesor might be difficult to filter and wash free of impurities, andparticle shape and size distribution might be different betweenpolymorphs).

As used herein and unless otherwise specified, the terms “solvate” and“solvated,” refer to a crystal form of a substance which containssolvent. The terms “hydrate” and “hydrated” refer to a solvate whereinthe solvent comprises water. “Polymorphs of solvates” refers to theexistence of more than one crystal form for a particular solvatecomposition. Similarly, “polymorphs of hydrates” refers to the existenceof more than one crystal form for a particular hydrate composition. Theterm “desolvated solvate,” as used herein, refers to a crystal form of asubstance which may be prepared by removing the solvent from a solvate.

As used herein and unless otherwise specified, the term “amorphous,”“amorphous form,” and related terms used herein, mean that thesubstance, component or product in question is not substantiallycrystalline as determined by X-ray diffraction. In particular, the term“amorphous form” describes a disordered solid form, i.e., a solid formlacking long range crystalline order. In certain embodiments, anamorphous form of a substance may be substantially free of otheramorphous forms and/or crystal forms. In other embodiments, an amorphousform of a substance may contain less than about 1%, 2%, 3%, 4%, 5%, 10%,15%, 20%, 25%, 30%, 35%, 40%, 45% or 50% of one or more other amorphousforms and/or crystal forms on a weight basis. In certain embodiments, anamorphous form of a substance may be physically and/or chemically pure.In certain embodiments, an amorphous form of a substance be about 99%,98%, 97%, 96%, 95%, 94%, 93%, 92%, 91% or 90% physically and/orchemically pure.

Techniques for characterizing crystal forms and amorphous forms include,but are not limited to, thermal gravimetric analysis (TGA), differentialscanning calorimetry (DSC), X-ray powder diffractometry (XRPD),single-crystal X-ray diffractometry, vibrational spectroscopy, e.g.,infrared (IR) and Raman spectroscopy, solid-state and solution nuclearmagnetic resonance (NMR) spectroscopy, optical microscopy, hot stageoptical microscopy, scanning electron microscopy (SEM), electroncrystallography and quantitative analysis, particle size analysis (PSA),surface area analysis, solubility measurements, dissolutionmeasurements, elemental analysis and Karl Fischer analysis.Characteristic unit cell parameters may be determined using one or moretechniques such as, but not limited to, X-ray diffraction and neutrondiffraction, including single-crystal diffraction and powderdiffraction. Techniques useful for analyzing powder diffraction datainclude profile refinement, such as Rietveld refinement, which may beused, e.g., to analyze diffraction peaks associated with a single phasein a sample comprising more than one solid phase. Other methods usefulfor analyzing powder diffraction data include unit cell indexing, whichallows one of skill in the art to determine unit cell parameters from asample comprising crystalline powder.

As used herein and unless otherwise specified, the terms “about” and“approximately,” when used in connection with a numeric value or a rangeof values which is provided to characterize a particular solid form,e.g., a specific temperature or temperature range, such as, e.g., thatdescribing a DSC or TGA thermal event, including, e.g., melting,dehydration, desolvation or glass transition events; a mass change, suchas, e.g., a mass change as a function of temperature or humidity; asolvent or water content, in terms of, e.g., mass or a percentage; or apeak position, such as, e.g., in analysis by IR or Raman spectroscopy orXRPD; indicate that the value or range of values may deviate to anextent deemed reasonable to one of ordinary skill in the art while stilldescribing the particular solid form. For example, in particularembodiments, the terms “about” and “approximately,” when used in thiscontext and unless otherwise specified, indicate that the numeric valueor range of values may vary within 25%, 20%, 15%, 10%, 9%, 8%, 7%, 6%,5%, 4%, 3%, 2%, 1.5%, 1%, 0.5%, or 0.25% of the recited value or rangeof values.

As used herein and unless otherwise specified, a sample comprising aparticular crystal form or amorphous form that is “substantially pure,”e.g., substantially free of other solid forms and/or of other chemicalcompounds, contains, in particular embodiments, less than about 25%,20%, 15%, 10%, 9%, 8%, 7%, 6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25% or0.1% percent by weight of one or more other solid forms and/or of otherchemical compounds.

As used herein and unless otherwise specified, a sample or compositionthat is “substantially free” of one or more other solid forms and/orother chemical compounds means that the composition contains, inparticular embodiments, less than about 25%, 20%, 15%, 10%, 9%, 8%, 7%,6%, 5%, 4%, 3%, 2%, 1%, 0.75%, 0.5%, 0.25% or 0.1% percent by weight ofone or more other solid forms and/or other chemical compounds.

As used herein, and unless otherwise specified, the terms “treat,”“treating” and “treatment” refer to the eradication or amelioration of adisease or disorder, or of one or more symptoms associated with thedisease or disorder. In certain embodiments, the terms refer tominimizing the spread or worsening of the disease or disorder resultingfrom the administration of one or more prophylactic or therapeuticagents to a patient with such a disease or disorder. In someembodiments, the terms refer to the administration of a compoundprovided herein, with or without other additional active agent, afterthe onset of symptoms of the particular disease.

As used herein, and unless otherwise specified, the terms “prevent,”“preventing” and “prevention” refer to the prevention of the onset,recurrence or spread of a disease or disorder, or of one or moresymptoms thereof. In certain embodiments, the terms refer to thetreatment with or administration of a compound provided herein, with orwithout other additional active compound, prior to the onset ofsymptoms, particularly to patients at risk of diseases or disordersprovided herein. The terms encompass the inhibition or reduction of asymptom of the particular disease. Patients with familial history of adisease in particular are candidates for preventive regimens in certainembodiments. In addition, patients who have a history of recurringsymptoms are also potential candidates for the prevention. In thisregard, the term “prevention” may be interchangeably used with the term“prophylactic treatment.”

As used herein, and unless otherwise specified, the terms “manage,”“managing” and “management” refer to preventing or slowing theprogression, spread or worsening of a disease or disorder, or of one ormore symptoms thereof. Often, the beneficial effects that a patientderives from a prophylactic and/or therapeutic agent do not result in acure of the disease or disorder. In this regard, the term “managing”encompasses treating a patient who had suffered from the particulardisease in an attempt to prevent or minimize the recurrence of thedisease.

As used herein, and unless otherwise specified, a “therapeuticallyeffective amount” of a compound is an amount sufficient to provide atherapeutic benefit in the treatment or management of a disease ordisorder, or to delay or minimize one or more symptoms associated withthe disease or disorder. A therapeutically effective amount of acompound means an amount of therapeutic agent, alone or in combinationwith other therapies, which provides a therapeutic benefit in thetreatment or management of the disease or disorder. The term“therapeutically effective amount” can encompass an amount that improvesoverall therapy, reduces or avoids symptoms or causes of disease ordisorder, or enhances the therapeutic efficacy of another therapeuticagent.

As used herein, and unless otherwise specified, a “prophylacticallyeffective amount” of a compound is an amount sufficient to prevent adisease or disorder, or prevent its recurrence. A prophylacticallyeffective amount of a compound means an amount of therapeutic agent,alone or in combination with other agents, which provides a prophylacticbenefit in the prevention of the disease. The term “prophylacticallyeffective amount” can encompass an amount that improves overallprophylaxis or enhances the prophylactic efficacy of anotherprophylactic agent.

The term “composition” as used herein is intended to encompass a productcomprising the specified ingredients (and in the specified amounts, ifindicated), as well as any product which results, directly orindirectly, from combination of the specified ingredients in thespecified amounts. By “pharmaceutically acceptable” it is meant that thediluent, excipient or carrier must be compatible with the otheringredients of the formulation and not deleterious to the recipientthereof

4. DETAILED DESCRIPTION OF THE INVENTION

This invention relates to stereomerically pure Compound A, which is the(+) enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione,substantially free of its (−) enantiomer, as well as novel methods ofusing, and compositions comprising, stereomerically pure Compound Aand/or solid forms comprising Compound A. For example, the presentinvention encompasses the in vitro and in vivo use of Compound A, andthe incorporation of Compound A into pharmaceutical compositions andsingle unit dosage forms useful in the treatment and prevention of avariety of diseases and disorders. Diseases and disorders which areameliorated by the reduction of levels of TNF-α or inhibition of PDE4are well known in the art and are described herein. Specific methods ofthe invention reduce or avoid the adverse effects associated withcompounds used as TNF-α inhibitor. Other specific methods of theinvention reduce or avoid the adverse effects associated with use ofracemic2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione.

Specific methods of the invention include methods of treating orpreventing diseases and disorders including, but not limited to, solidtumors, blood-borne tumors and inflammatory diseases.

Pharmaceutical and dosage forms of the invention, which compriseCompound A or a pharmaceutically acceptable polymorph, prodrug,clathrate, solvate or hydrate thereof (wherein particular embodimentsencompass solid forms comprising Compound A as described herein) can beused in the methods of the invention.

Without being limited by theory, it is believed that Compound A,including solid forms comprising Compound A, can inhibit TNF-αproduction. Consequently, a first embodiment of the invention relates toa method of inhibiting TNF-α production which comprises contacting acell exhibiting abnormal TNF-α production with an effective amount ofstereomerically pure Compound A or a pharmaceutically acceptableprodrug, metabolite, polymorph, solvate, hydrate, or clathrate thereof(wherein particular embodiments encompass solid forms comprisingCompound A as described herein). In a particular embodiment, theinvention relates to a method of inhibiting TNF-α production whichcomprises contacting a mammalian cell exhibiting abnormal TNF-αproduction with an effective amount of stereomerically pure Compound Aor a pharmaceutically acceptable prodrug, metabolite, polymorph,solvate, hydrate, or clathrate thereof (wherein particular embodimentsencompass solid forms comprising Compound A as described herein).

The invention also relates to a method of treating, preventing ormanaging disorders ameliorated by the reduction of levels of TNF-α in apatient which comprises administering to a patient in need of suchtreatment or prevention a therapeutically or prophylactically effectiveamount of stereomerically pure Compound A or a pharmaceuticallyacceptable prodrug, metabolite, polymorph, solvate, hydrate, orclathrate thereof (wherein particular embodiments encompass solid formscomprising Compound A as described herein). In particular embodiments,diseases or disorders ameliorated by the inhibition of TNF-α productionin mammals include, but are not limited to: HIV; hepatitis; adultrespiratory distress syndrome; bone resorption diseases; chronicobstructive pulmonary diseases; chronic pulmonary inflammatory diseases;asthma; dermatitis; cystic fibrosis; septic shock; sepsis; endotoxicshock; hemodynamic shock; sepsis syndrome; post ischemic reperfusioninjury; meningitis; psoriasis; fibrotic disease; cachexia; graftrejection; auto immune disease; rheumatoid spondylitis; arthriticconditions, such as psoriatic arthritis, rheumatoid arthritis andosteoarthritis; osteoporosis; Crohn's disease; ulcerative colitis;inflammatory bowel disease; multiple sclerosis; systemic lupuserythematosus; cutaneous lupus erythematosus; pulmonary sarcoidosis;erythema nodosum leprosum (ENL) in leprosy; radiation damage; asthma;and hyperoxic alveolar injury. Such disorders further include, but arenot limited to, cancers, including, but not limited to cancer of thehead, thyroid, neck, eye, skin, mouth, throat, esophagus, chest, bone,blood, bone marrow, lung, colon, sigmoid, rectum, stomach, prostate,breast, ovaries, kidney, liver, pancreas, brain, intestine, heart,adrenal, subcutaneous tissue, lymph nodes, heart, and combinationsthereof. Specific cancers that can be treated by this method aremultiple myeloma, malignant melanoma, malignant glioma, leukemia andsolid tumors.

A further embodiment of the invention relates to a method of treating orpreventing cancer, including but not limited to, solid tumor,blood-borne tumor, leukemias, and in particular, multiple myeloma in apatient which comprises administering to a patient in need of suchtreatment or prevention a therapeutically effective amount ofstereomerically pure Compound A or a pharmaceutically acceptableprodrug, metabolite, polymorph, solvate, hydrate, or clathrate thereof(wherein particular embodiments encompass solid forms comprisingCompound A as described herein); in particular wherein the patient is amammal.

In another embodiment, the invention relates to a method of inhibitingPDE4 which comprises contacting PDE4 in a cell (e.g. a mammalian cell)with an effective amount of stereomerically pure Compound A or apharmaceutically acceptable prodrug, metabolite, polymorph, solvate,hydrate, or clathrate thereof (wherein particular embodiments encompasssolid forms comprising Compound A as described herein).

A further embodiment of the invention relates to a method of treating orpreventing diseases or disorders ameliorated by the inhibition of PDE4in a patient which comprises administering to a patient in need of suchtreatment or prevention a therapeutically or prophylactically effectiveamount of stereomerically pure Compound A or a pharmaceuticallyacceptable prodrug, metabolite, polymorph, solvate, hydrate, orclathrate thereof (wherein particular embodiments encompass solid formscomprising Compound A as described herein). Disorders ameliorated by theinhibition of PDE4 include, but are not limited to, asthma, inflammation(e.g., inflammation due to reperfusion), chronic or acute obstructivepulmonary diseases, chronic or acute pulmonary inflammatory diseases,inflammatory bowel disease, Crohn's Disease, Behcet's Disease, orcolitis.

In another embodiment, the invention relates to a method of controllingcAMP levels in a cell which comprises contacting a cell with aneffective amount of stereomerically pure Compound A or apharmaceutically acceptable prodrug, metabolite, polymorph, solvate,hydrate, or clathrate thereof (wherein particular embodiments encompasssolid forms comprising Compound A as described herein). As used hereinthe term “controlling cAMP levels” includes preventing or reducing therate of the breakdown of Adenosine 3′,5′-cyclic monophosphatc (cAMP) ina cell or increasing the amount of Adenosine 3′,5′-cyclic monophosphatepresent in a cell, preferably a mammalian cell, more preferably a humancell. In a particular method, the rate of cAMP breakdown is reduced byabout 10, 25, 50, 100, 200, or 500 percent as compared to the rate incomparable cells which have not been contacted with a compound of theinvention.

A further embodiment of the invention relates to a method of treating orpreventing depression, asthma, inflammation (e.g., contact dermatitis,atopic dermatitis, psoriasis, rheumatoid arthritis, osteoarthritis,inflammatory skin disease, inflammation due to reperfusion), chronic oracute obstructive pulmonary diseases, chronic or pulmonary inflammatorydiseases, inflammatory bowel disease, Crohn's Disease, Behcet's Diseaseor colitis in a patient which comprises administering to a patient inneed of such treatment or prevention a therapeutically orprophylactically effective amount of stereomerically pure Compound A ora pharmaceutically acceptable prodrug, metabolite, polymorph, solvate,hydrate, or clathrate thereof (wherein particular embodiments encompasssolid forms comprising Compound A as described herein); in particularwherein the patient is a mammal.

A separate embodiment of the invention encompasses methods of treatingor preventing myelodysplastic syndrome (MDS) which comprisesadministering to a patient in need of such treatment or prevention atherapeutically or prophylactically effective amount of stereomericallypure Compound A or a pharmaceutically acceptable solvate, hydrate,stereoisomer, clathrate, or prodrug thereof (wherein particularembodiments encompass solid forms comprising Compound A as describedherein). MDS refers to a diverse group of hematopoietic stem celldisorders. MDS is characterized by a cellular marrow with impairedmorphology and maturation (dysmyelopoiesis), peripheral bloodcytopenias, and a variable risk of progression to acute leukemia,resulting from ineffective blood cell production. See The Merck Manual953 (17th ed. 1999) and List et al., 1990, J. Clin. Oncol. 8:1424.

A separate embodiment of the invention encompasses methods of treatingor preventing myeloproliferative disease (MPD) which comprisesadministering to a patient in need of such treatment or prevention atherapeutically or prophylactically effective amount of stereomericallypure Compound A or a pharmaceutically acceptable solvate, hydrate,stereoisomer, clathrate, or prodrug thereof (wherein particularembodiments encompass solid forms comprising Compound A as describedherein). Myeloproliferative disease (MPD) refers to a group of disorderscharacterized by clonal abnormalities of the hematopoietic stem cell.See e.g., Current Medical Diagnosis & Treatment, pp. 499 (37th ed.,Tierney et al., ed., Appleton & Lange, 1998).

The invention also encompasses a method of treating, preventing ormanaging pain, including, but not limited to, complex regional painsyndrome, which comprises administering to a patient in need of suchtreatment, prevention or management a therapeutically orprophylactically effective amount of a stereomerically pure Compound Aor a pharmaceutically acceptable solvate, hydrate, stereoisomer,clathrate, or prodrug thereof (wherein particular embodiments encompasssolid forms comprising Compound A as described herein). In a specificembodiment, the administration is before, during or after surgery orphysical therapy directed at reducing or avoiding a symptom of complexregional pain syndrome in the patient.

In particular methods of the invention, stereomerically pure Compound Aor a pharmaceutically acceptable polymorph, prodrug, solvate, hydrate,or clathrate thereof (wherein particular embodiments encompass solidforms comprising Compound A as described herein), is adjunctivelyadministered with at least one additional therapeutic agent. Examples ofadditional therapeutic agents include, but are not limited to,anti-cancer drugs, anti-inflammatories, antihistamines anddecongestants.

4.1. Solid Forms Comprising Compound A

Certain embodiments herein provide solid forms comprising Compound A,which has the chemical structure shown above. Racemic2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneis readily prepared using the methods in U.S. Pat. No. 6,020,358, whichis incorporated herein by reference. Compound A, which is the (+)enantiomer of2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione,can be prepared according to any method apparent to those of skill inthe art, including the methods described in U.S. Pat. No. 6,962,940,which is incorporated herein by reference.

Solid forms comprising Compound A include single-component andmultiple-component forms, including crystal forms and amorphous forms,and including, but not limited to, polymorphs, solvates, hydrates,co-crystals and clathrates. Particular embodiments herein providesingle-component amorphous solid forms of Compound A. Particularembodiments herein provide single-component crystalline solid forms ofCompound A. Particular embodiments herein provide multiple-componentamorphous forms comprising Compound A. Particular embodiments hereinprovide multiple-component crystalline solid forms comprising CompoundA. Multiple-component solid forms provided herein include solid formswhich may be described by the terms salt, co-crystal, hydrate, solvate,clathrate and/or polymorph, and include solid forms which may bedescribed by one or more of these terms.

Solid forms comprising Compound A can be prepared by the methodsdescribed herein, including the methods described in the Examples below,or by techniques known in the art, including heating, cooling, freezedrying, lyophilization, quench cooling the melt, rapid solventevaporation, slow solvent evaporation, solvent recrystallization,antisolvent addition, slurry recrystallization, crystallization from themelt, desolvation, recrystallization in confined spaces such as, e.g.,in nanopores or capillaries, recrystallization on surfaces or templatessuch as, e.g., on polymers, recrystallization in the presence ofadditives, such as, e.g., co-crystal counter-molecules, desolvation,dehydration, rapid cooling, slow cooling, exposure to solvent and/orwater, drying, including, e.g., vacuum drying, vapor diffusion,sublimation, grinding (including, e.g., cryo-grinding, solvent-dropgrinding or liquid assisted grinding), microwave-induced precipitation,sonication-induced precipitation, laser-induced precipitation andprecipitation from a supercritical fluid. The particle size of theresulting solid forms, which can vary, (e.g., from nanometer dimensionsto millimeter dimensions), can be controlled, e.g., by varyingcrystallization conditions, such as, e.g., the rate of crystallizationand/or the crystallization solvent system, or by particle-size reductiontechniques, e.g., grinding, milling, micronizing or sonication.

While not intending to be bound by any particular theory, certain solidforms are characterized by physical properties, e.g., stability,solubility and dissolution rate, appropriate for pharmaceutical andtherapeutic dosage forms. Moreover, while not wishing to be bound by anyparticular theory, certain solid forms are characterized by physicalproperties (e.g., density, compressibility, hardness, morphology,cleavage, stickiness, solubility, water uptake, electrical properties,thermal behavior, solid-state reactivity, physical stability, andchemical stability) affecting particular processes (e.g., yield,filtration, washing, drying, milling, mixing, tableting, flowability,dissolution, formulation, and lyophilization) which make certain solidforms suitable for the manufacture of a solid dosage form. Suchproperties can be determined using particular analytical chemicaltechniques, including solid-state analytical techniques (e.g., X-raydiffraction, microscopy, spectroscopy and thermal analysis), asdescribed herein and known in the art.

Certain embodiments herein provide compositions comprising one or moreof the solid forms. Certain embodiments provide compositions of one ormore solid forms in combination with other active ingredients. Certainembodiments provide methods of using these compositions in thetreatment, prevention or management of diseases and disorders including,but not limited to, the diseases and disorders provided herein.

In addition to solid forms comprising Compound A, provided herein aresolid forms comprising prodrugs of Compound A.

Solid forms provided herein may also comprise unnatural proportions ofatomic isotopes at one or more of the atoms in Compound A. For example,the compound may be radiolabeled with radioactive isotopes, such as forexample tritium (³H), iodine-125 (¹²⁵I) sulfur-35 (³⁵S), or carbon-14(¹⁴C). Radiolabeled compounds are useful as therapeutic agents, e.g.,cancer therapeutic agents, research reagents, e.g., binding assayreagents, and diagnostic agents, e.g., in vivo imaging agents. Allisotopic variations of Compound A, whether radioactive or not, areintended to be encompassed within the scope of the embodiments providedherein.

4.1.1. Form A of Compound A

Certain embodiments herein provide the Form A crystal form of CompoundA. In certain embodiments, Form A of Compound A can be obtained fromvarious solvents, including, but not limited to, solvent systemscomprising acetone, ethanol, and mixtures thereof. In certainembodiments, Form A can be obtained using a fast cooling crystallizationprocess.

In certain embodiments, Form A of Compound A may be characterized byX-ray powder diffraction analysis. A representative XRPD pattern of FormA of Compound A is provided in FIG. 1. In certain embodiments, Form A ofCompound A is characterized by XRPD peaks located at one, two, three,four, five, six, seven, eight, nine, ten, eleven or twelve of thefollowing approximate positions: 8.1, 14.4, 15.2, 17.4, 18.4, 19.2,20.5, 22.8, 23.2 23.6, 24.5, 25.1 degrees 2θ. In certain embodiments,Form A of Compound A is characterized by an XRPD pattern which matchesthe pattern exhibited in FIG. 1. In certain embodiments, Form A ofCompound A is characterized by an XRPD pattern having 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or25 peaks matching peaks in the representative Form A pattern providedherein.

In certain embodiments, Form A of Compound A may be characterized bythermal analysis. A representative DSC plot for Form A of Compound A isshown in FIG. 2. In certain embodiments, Form A is characterized by aDSC plot comprising an endothermic event with an onset temperature ofabout 145° C. In certain embodiments, Form A is characterized by a DSCplot further comprising an endothermic event with an onset temperatureof about 155° C. A representative TGA plot for Form A of Compound A isshown in FIG. 3. In certain embodiments, Form A is characterized by aTGA plot comprising a mass loss of less than about 1%, e.g., about0.05%, of the total mass of the sample upon heating from about 25° C. toabout 140° C. In certain embodiments, Form A of Compound A does notcontain substantial amounts of either water or other solvent in thecrystal lattice. In certain embodiments, Form A is unsolvated. Incertain embodiments, Form A is anhydrous.

In certain embodiments, Form A of Compound A may be characterized bymoisture sorption analysis. A representative moisture sorption isothermplot is shown in FIG. 4. In certain embodiments, when the relativehumidity (“RH”) is increased from about 0% to about 95% RH, Form Aexhibits a mass change of less than about 1%, e.g., about 0.4%, of thestarting mass of the sample. In certain embodiments, mass gained uponadsorption is lost when the RH is decreased back to about 0% RH.Accordingly, in certain embodiments, Form A is substantiallynonhygroscopic. In certain embodiments, the XRPD pattern of the Form Amaterial is substantially unchanged following the adsorption/desorptionanalysis. In certain embodiments, Form A is stable with respect tohumidity.

In certain embodiments, Form A of Compound A may be characterized by itsstability profile. In certain embodiments, Form A material is stable,e.g., its XRPD pattern remains substantially unchanged, upon exposure toelevated temperature, upon exposure to elevated humidity, upon exposureto one or more solvents, and/or upon compression. In certainembodiments, for example, Form A is stable following exposure to anenvironment of about 40° C. and about 75% RH environment for about fourweeks. In certain embodiments, Form A is stable following exposure toone or more solvent systems comprising, e.g., ethanol, water and/orheptane, at about 40° C. for at least about four weeks. In certainembodiments, Form A converts to Form C of Compound A upon exposure to asolvent including, but not limited to, toluene for four weeks. Incertain embodiments, Form A is stable upon compression at about 2000 psipressure for about one minute.

In certain embodiments, Form A of Compound A may be characterized byparticle analysis. In certain embodiments, Form A is characterized as awhite powder. In certain embodiments, a sample of Form A comprisesparticles having a plate-like morphology. In certain embodiments, asample of Form A comprises particles with a D₉₀ of less than about 18(As used herein, the D₉₀ value represents the 90th percentile of theparticle size distribution as measured by length; i.e., 90% of theparticles have a length of this value or less).

Certain embodiments herein provide Form A of Compound A which issubstantially pure. Certain embodiments herein provide Form A ofCompound A which is substantially free of other solid forms comprisingCompound A including, e.g., Forms B, C, D, E, F, G and/or an amorphoussolid form comprising Compound A as provided herein. Certain embodimentsherein provide Form A as a mixture of solid forms comprising Compound A,including, e.g., a mixture comprising one or more of the following:Forms B, C, D, E, F, G and an amorphous solid form comprising Compound Aas provided herein.

4.1.2. Form B of Compound A

Certain embodiments herein provide the Form B crystal form of CompoundA. In certain embodiments, Form B of Compound A can be obtained fromvarious solvents, including, but not limited to, solvent systemscomprising 2-propanol, acetone, acetonitrile, ethanol, ethyl acetate,heptane, methanol, methyl ethyl ketone, methyl t-butyl ether, methylenechloride, n-butanol, n-butyl acetate, tetrahydrofuran, toluene, waterand mixtures comprising two or more thereof. For example, in certainembodiments, Form B can be obtained by crystallization from a solventsystem comprising 1:1 ethanol:water, e.g., by a process comprisingevaporation of the 1:1 ethanol:water solvent system at about 25° C.,followed by isolation of Form B. For example, in certain embodiments,Form B can be obtained by crystallization from a solvent systemcomprising 1:1 acetone:ethanol, e.g., by a process comprising slurryinga solid form comprising Compound A in 1:1 acetone:ethanol at about 25°C. for about 2 days, followed by isolation of Form B.

In certain embodiments, Form B of Compound A may be characterized byX-ray powder diffraction analysis. A representative XRPD pattern of FormB of Compound A is provided in FIG. 5. In certain embodiments, Form B ofCompound A is characterized by XRPD peaks located at one, two, three,four, five, six, seven, eight, nine, ten, eleven or twelve of thefollowing approximate positions: 10.1, 12.4, 13.5, 15.7, 16.3, 18.1,20.7, 22.5, 24.7, 26.2, 26.9, 29.1 degrees 2θ. In certain embodiments,Form B of Compound A is characterized by an XRPD pattern which matchesthe pattern exhibited in FIG. 5. In certain embodiments, Form B ofCompound A is characterized by an XRPD pattern having 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or25 peaks matching peaks in the representative Form B pattern providedherein.

In certain embodiments, Form B of Compound A may be characterized bythermal analysis. A representative DSC plot for Form B of Compound A isshown in FIG. 6. In certain embodiments, Form B is characterized by aDSC plot comprising an endothermic event with an onset temperature ofabout 154° C. A representative TGA plot for Form B of Compound A isshown in FIG. 7. In certain embodiments, Form B is characterized by aTGA plot comprising a mass loss of less than about 1%, e.g., about0.25%, of the total mass of the sample upon heating from about 25° C. toabout 140° C. In certain embodiments, Form B of Compound A does notcontain substantial amounts of either water or other solvent in thecrystal lattice. In certain embodiments, Form B is anhydrous. In certainembodiments, Form B is unsolvated.

In certain embodiments, Form B of Compound A may be characterized bymoisture sorption analysis. A representative moisture sorption isothermplot is shown in FIG. 8. In certain embodiments, when the RH isincreased from about 0% to about 95% RH, Form B exhibits a mass changeof less than about 1%, e.g., about 0.6%, of the starting mass of thesample. In certain embodiments, mass gained upon adsorption is lost whenthe RH is decreased back to about 0% RH. In certain embodiments, Form Bis substantially nonhygroscopic. In certain embodiments, the XRPDpattern of Form B material is substantially unchanged following theadsorption/desorption analysis. In certain embodiments, Form B is stablewith respect to humidity.

In certain embodiments, Form B of Compound A may be characterized by itsstability profile. In certain embodiments, Form B material is stable,e.g., its XRPD pattern remains substantially unchanged, upon exposure toelevated temperature, upon exposure to elevated humidity, upon exposureto one or more solvents, and/or upon compression. In certainembodiments, for example, Form B is stable following exposure to anenvironment of about 40° C. and about 75% RH environment for about fourweeks. In certain embodiments, Form B is stable following exposure to asolvent system comprising, e.g., ethanol, water or heptane, at about 40°C. for at least about four weeks. In certain embodiments, Form Bconverts to Form C of Compound A upon exposure to a solvent systemcomprising, e.g., toluene for about four weeks. In certain embodiments,Form B is stable following compression at about 2000 psi pressure forabout one minute.

In certain embodiments, Form B of Compound A may be characterized byparticle analysis. In certain embodiments, Form B is characterized as awhite powder. In certain embodiments, a sample of Form B comprisesparticles having a flake-like morphology. In certain embodiments, asample of Form B comprises particles with a D₉₀ of less than about 12μm.

Certain embodiments herein provide Form B of Compound A which issubstantially pure. Certain embodiments herein provide Form B ofCompound A which is substantially free of other solid forms comprisingCompound A including, e.g., Forms A, C, D, E, F, G and/or an amorphoussolid form comprising Compound A as provided herein. Certain embodimentsherein provide Form B as a mixture of solid forms comprising Compound A,including, e.g., a mixture comprising one or more of the following:Forms A, C, D, E, F, G and an amorphous solid form comprising Compound Aas provided herein.

4.1.3. Form C of Compound A

Certain embodiments herein provide the Form C crystal form of CompoundA. In certain embodiments, Form C of Compound A can be obtained fromvarious solvent systems, including, but not limited to, solvent systemscomprising acetone, acetonitrile, ethanol, heptane, methanol, methylethyl ketone, tetrahydrofuran, toluene, water, and mixtures comprisingtwo or more thereof. For example, in certain embodiments, Form C can beobtained by crystallization from a solvent system comprising toluene,e.g., by a process comprising the use of toluene as an anti-solvent,followed by isolation of Form C.

In certain embodiments, Form C of Compound A may be characterized byX-ray powder diffraction analysis. A representative XRPD pattern of FormC of Compound A is provided in FIG. 9. In certain embodiments, Form C ofCompound A is characterized by XRPD peaks located at one, two, three,four, five, six, seven, eight, nine, ten, eleven or twelve of thefollowing approximate positions: 7.5, 11.3, 15.3, 16.4, 17.8, 21.4,22.6, 23.5, 24.8, 25.5, 26.4, 27.6 degrees 2θ. In certain embodiments,Form C of Compound A is characterized by an XRPD pattern which matchesthe pattern exhibited in FIG. 9. In certain embodiments, Form C ofCompound A is characterized by an XRPD pattern having 1, 2, 3, 4, 5, 6,7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or25 peaks matching peaks in the representative Form C pattern providedherein.

In certain embodiments, Form C of Compound A may be characterized bythermal analysis. A representative DSC plot for Form C of Compound A isshown in FIG. 10. In certain embodiments, Form C is characterized by aDSC plot comprising an endothermic event with an onset temperature ofabout 138° C. In certain embodiments, a characteristic Form C DSC plotfurther comprises one or more additional events, such as, e.g., anendothermic event with an onset temperature of about 166° C. Arepresentative TGA plot for Form C of Compound A is shown in FIG. 11. Incertain embodiments, Form C is characterized by a TGA plot comprising amass loss of less than about 10%, e.g., about 5.9%, of the total mass ofthe sample upon heating from about 25° C. to about 140° C. In certainembodiments, the TGA mass loss event comprises the loss of the solventtoluene, as indicated, e.g., by TG-IR analysis. In certain embodiments,Form C of Compound A is solvated. In certain embodiments, Form C is atoluene solvate. In certain embodiments, the crystal lattice of Form Ccomprises about three molar equivalents of toluene per mole of CompoundA.

In certain embodiments, Form C of Compound A may be characterized bymoisture sorption analysis. A representative moisture sorption isothermplot is shown in FIG. 12. In certain embodiments, when the RH isincreased from about 0% to about 95% RH, Form C exhibits a mass changeof less than about 1%, e.g., about 0.5%, of the starting mass of thesample. In certain embodiments, mass gained upon adsorption is lost whenthe RH is decreased back to about 0% RH. In certain embodiments, Form Cis substantially nonhygroscopic. In certain embodiments, the XRPDpattern of Form C material is substantially unchanged following theadsorption/desorption analysis. In certain embodiments, Form C is stablewith respect to humidity.

In certain embodiments, Form C of Compound A may be characterized by itsstability profile. In certain embodiments, Form C material is stable,e.g., its XRPD pattern remains substantially unchanged, upon exposure toelevated temperature, upon exposure to elevated humidity, upon exposureto one or more solvents, and/or upon compression. In certainembodiments, for example, Form C is stable following exposure to anenvironment of about 40° C. and about 75% RH environment for about fourweeks. In certain embodiments, Form C is stable following exposure to asolvent system comprising, e.g., ethanol, water, heptane or toluene, atabout 40° C. for at least about four weeks. In certain embodiments, FormC is stable following compression at about 2000 psi pressure for aboutone minute.

In certain embodiments, Form C of Compound A may be characterized byparticle analysis. In certain embodiments, Form C is characterized as awhite powder. In certain embodiments, a sample of Form C comprisesparticles having a plate-like morphology. In certain embodiments, asample of Form C comprises particles with a D₉₀ of less than about 12μm.

Certain embodiments herein provide Form C of Compound A which issubstantially pure. Certain embodiments herein provide Form C ofCompound A which is substantially free of other solid forms comprisingCompound A including, e.g., Forms A, B, D, E, F, G and/or an amorphoussolid form comprising Compound A as provided herein. Certain embodimentsherein provide Form C as a mixture of solid forms comprising Compound A,including, e.g., a mixture comprising one or more of the following:Forms A, B, D, E, F, G and an amorphous solid form comprising Compound Aas provided herein.

4.1.4. Form D of Compound A

Certain embodiments herein provide the Form D crystal form of CompoundA. In certain embodiments, Form D of Compound A can be obtained fromvarious solvents, including, but not limited to, solvent systemscomprising methylene chloride. For example, in certain embodiments, FormD can be obtained by crystallization from a solvent system comprisingmethylene chloride, e.g., by a process comprising the evaporation ofmethylene chloride, followed by isolation of Form D.

In certain embodiments, Form D of Compound A may be characterized byX-ray powder diffraction analysis. A representative XRPD pattern of FormD of Compound A is provided in FIG. 13. In certain embodiments, Form Dof Compound A is characterized by XRPD peaks located at one, two, three,four, five, six, seven, eight, nine, ten, eleven or twelve of thefollowing approximate positions: 7.5, 9.6, 11.3, 13.9, 16.3, 17.7, 20.5,23.2, 24.6, 25.2, 26.0, 28.8 degrees 2θ. In certain embodiments, Form Dof Compound A is characterized by an XRPD pattern which matches thepattern exhibited in FIG. 13. In certain embodiments, Form D of CompoundA is characterized by an XRPD pattern having 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 peaksmatching peaks in the representative Form D pattern provided herein.

In certain embodiments, Form D of Compound A may be characterized bythermal analysis. A representative DSC plot for Form D of Compound A isshown in FIG. 14. In certain embodiments, Form D is characterized by aDSC plot comprising an endothermic event with an onset temperature ofabout 100° C. A representative TGA plot for Form D of Compound A isshown in FIG. 15. In certain embodiments, Form D is characterized by aTGA plot comprising a mass loss of less than about 10%, e.g., about6.5%, of the total mass of the sample upon heating from about 25° C. toabout 110° C. In certain embodiments, the TGA mass loss event comprisesthe loss of the solvent methylene chloride (i.e. dichloromethane), asindicated, e.g., by TG-IR analysis. In certain embodiments, Form D ofCompound A is solvated. In certain embodiments, Form D is a methylenechloride solvate. In certain embodiments, the crystal lattice of Form Dcomprises about 2.5 molar equivalents of methylene chloride per mole ofCompound A.

In certain embodiments, Form D of Compound A may be characterized bymoisture sorption analysis. A representative moisture sorption isothermplot is shown in FIG. 16. In certain embodiments, when the RH isincreased from about 0% to about 95% RH, Form D exhibits a mass changeof less than about 3%, e.g., about 1.5%, of the starting mass of thesample. In certain embodiments, mass gained upon adsorption is lost whenthe RH is decreased back to about 0% RH. Accordingly, in certainembodiments, Form D is slightly hygroscopic. In certain embodiments, theXRPD pattern of Form D material is substantially unchanged following theadsorption/desorption analysis. In certain embodiments, Form D is stablewith respect to humidity.

In certain embodiments, Form D of Compound A may be characterized by itsstability profile. In certain embodiments, Form D material is stable,e.g., its XRPD pattern remains substantially unchanged, uponcompression. For example, in certain embodiments, Form D is stablefollowing compression at about 2000 psi pressure for about one minute.In certain embodiments, Form D is stable following exposure to anenvironment of about 40° C. and about 75% RH environment for about fourweeks, although, in certain embodiments, the resulting peak intensity ofthe Form D XRPD pattern is reduced. In certain embodiments, thisreduction in XRPD peak intensity results from the formation of amorphousmaterial comprising Compound A. In certain embodiments, Form D convertsto Form B of Compound A upon exposure to a solvent system comprising,e.g., heptane, ethanol and/or water at about 40° C. for about fourweeks. In certain embodiments, Form D converts to Form C of Compound Aupon exposure to a solvent system comprising toluene at about 40° C. forabout four weeks.

In certain embodiments, Form D of Compound A may be characterized byparticle analysis. In certain embodiments, Form D is characterized as awhite powder. In certain embodiments, a sample of Form D comprisesparticles having a flake-like morphology. In certain embodiments, asample of Form D comprises particles with a D₉₀ of less than about 18μm.

Certain embodiments herein provide Form D of Compound A which issubstantially pure. Certain embodiments herein provide Form D ofCompound A which is substantially free of other solid forms comprisingCompound A including, e.g., Forms A, B, C, E, F, G and/or an amorphoussolid form comprising Compound A as provided herein. Certain embodimentsherein provide Form D as a mixture of solid forms comprising Compound A,including, e.g., a mixture comprising one or more of the following:Forms A, B, C, E, F, G and an amorphous solid form comprising Compound Aas provided herein.

4.1.5. Form E of Compound A

Certain embodiments herein provide the Form E crystal form of CompoundA. In certain embodiments, Form E of Compound A can be obtained fromvarious solvents, including, but not limited to, solvent systemscomprising acetone, acetonitrile, heptane, methylene chloride, andmixtures comprising two or more thereof. For example, in certainembodiments, Form E can be obtained by crystallization from a solventsystem comprising acetonitrile, e.g., by a process comprising theevaporation of acetonitrile, followed by isolation of Form E.

In certain embodiments, Form E of Compound A may be characterized byX-ray powder diffraction analysis. A representative XRPD pattern of FormE of Compound A is provided in FIG. 17. In certain embodiments, Form Eof Compound A is characterized by XRPD peaks located at one, two, three,four, five, six, seven, eight, nine, ten, eleven or twelve of thefollowing approximate positions: 7.6, 9.2, 11.4, 15.5, 16.5, 17.9, 19.6,20.5, 21.6, 22.8, 23.8, 26.6 degrees 2θ. In certain embodiments, Form Eof Compound A is characterized by an XRPD pattern which matches thepattern exhibited in FIG. 17. In certain embodiments, Form E of CompoundA is characterized by an XRPD pattern having 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 peaksmatching peaks in the representative Form E pattern provided herein.

In certain embodiments, Form E of Compound A may be characterized bythermal analysis. A representative DSC plot for Form E of Compound A isshown in FIG. 18. In certain embodiments, Form E is characterized by aDSC plot comprising an endothermic event with an onset temperature ofabout 95° C. A representative TGA plot for Form E of Compound A is shownin FIG. 19. In certain embodiments, Form E is characterized by a TGAplot comprising a mass loss of less than about 8%, e.g., about 4.0%, ofthe total mass of the sample upon heating from about 25° C. to about110° C. In certain embodiments, the TGA mass loss event comprises theloss of the solvent acetonitrile, as indicated, e.g., by TG-IR analysis.In certain embodiments, Form E of Compound A is solvated. In certainembodiments, Form E is an acetonitrile solvate. In certain embodiments,the crystal lattice of Form E comprises about 2.5 molar equivalents ofacetonitrile per mole of Compound A.

In certain embodiments, Form E of Compound A may be characterized bymoisture sorption analysis. A representative moisture sorption isothermplot is shown in FIG. 20. In certain embodiments, when the RH isincreased from about 0% to about 95% RH, Form E exhibits a mass changeof less than about 10%, e.g., about 5.1%, of the starting mass of thesample. In certain embodiments, mass gained upon adsorption is lost whenthe RH is decreased back to about 0% RH. In certain embodiments, Form Eis hygroscopic. In certain embodiments, the XRPD pattern of Form Ematerial is substantially unchanged following the adsorption/desorptionanalysis. In certain embodiments, Form E is stable with respect tohumidity.

In certain embodiments, Form E of Compound A may be characterized by itsstability profile. In certain embodiments, Form E material is stable,e.g., its XRPD pattern remains substantially unchanged, uponcompression. For example, in certain embodiments, Form E is stablefollowing compression at about 2000 psi pressure for about one minute.

In certain embodiments, Form E of Compound A may be characterized byparticle analysis. In certain embodiments, Form E is characterized as awhite powder. In certain embodiments, a sample of Form E comprisesparticles having a flake-like morphology. In certain embodiments, asample of Form E comprises particles with a D₉₀ of less than about 18μm.

Certain embodiments herein provide Form E of Compound A which issubstantially pure. Certain embodiments herein provide Form E ofCompound A which is substantially free of other solid forms comprisingCompound A including, e.g., Forms A, B, C, D, F, G and/or an amorphoussolid form comprising Compound A as provided herein. Certain embodimentsherein provide Form E as a mixture of solid forms comprising Compound A,including, e.g., a mixture comprising one or more of the following:Forms A, B, C, D, F, G and an amorphous solid form comprising Compound Aas provided herein.

4.1.6. Form F of Compound A

Certain embodiments herein provide the Form F crystal form of CompoundA. In certain embodiments, Form F of Compound A can be obtained fromvarious solvents, including, but not limited to, solvent systemscomprising acetone, ethanol, water, and mixtures comprising two or morethereof. For example, in certain embodiments, Form F can be obtained bycrystallization from a solvent system comprising ethanol and/or water,e.g., by a process comprising contacting a solid form comprisingCompound A with a solvent system comprising ethanol and/or water,followed by isolation of Form F.

In certain embodiments, Form F of Compound A may be characterized byX-ray powder diffraction analysis. A representative XRPD pattern of FormF of Compound A is provided in FIG. 21. In certain embodiments, Form Fof Compound A is characterized by XRPD peaks located at one, two, three,four, five, six, seven, eight, nine, ten, eleven or twelve of thefollowing approximate positions: 8.1, 8.6, 15.6, 17.3, 19.3, 21.4, 22.8,24.6, 25.4, 25.9, 26.6, 27.7 degrees 2θ. In certain embodiments, Form Fof Compound A is characterized by an XRPD pattern which matches thepattern exhibited in FIG. 21. In certain embodiments, Form F of CompoundA is characterized by an XRPD pattern having 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 peaksmatching peaks in the representative Form F pattern provided herein.

In certain embodiments, Form F of Compound A may be characterized bythermal analysis. A representative DSC plot for Form F of Compound A isshown in FIG. 22. In certain embodiments, Form F is characterized by aDSC plot comprising an endothermic event with an onset temperature ofabout 145° C. A representative TGA plot for Form F of Compound A isshown in FIG. 23. In certain embodiments, Form F is characterized by aTGA plot comprising a mass loss of less than about 1%, e.g., about 0.1%,of the total mass of the sample upon heating from about 25° C. to about180° C. In certain embodiments, Form F of Compound A does not containsubstantial amounts of either water or other solvent in the crystallattice. In certain embodiments, Form F is unsolvated. In certainembodiments, Form F is anhydrous.

In certain embodiments, Form F of Compound A may be characterized bymoisture sorption analysis. A representative moisture sorption isothermplot is shown in FIG. 24. In certain embodiments, when the RH isincreased from about 0% to about 95% RH, Form F exhibits a mass changeof less than about 1%, e.g., about 0.2%, of the starting mass of thesample. In certain embodiments, mass gained upon adsorption is lost whenthe RH is decreased back to about 0% RH. In certain embodiments, Form Fis substantially nonhygroscopic. In certain embodiments, the XRPDpattern of Form F material is substantially unchanged following theadsorption/desorption analysis. In certain embodiments, Form F is stablewith respect to humidity.

In certain embodiments, Form F of Compound A may be characterized by itsstability profile. In certain embodiments, Form F material is stable,e.g., its XRPD pattern remains substantially unchanged, uponcompression. For example, in certain embodiments, Form F is stablefollowing compression at about 2000 psi pressure for about one minute.In certain embodiments, Form F is stable following exposure to a solventsystem comprising, e.g., ethanol, acetone or mixtures thereof, for abouttwo days at about 25° C.

In certain embodiments, Form F of Compound A may be characterized byparticle analysis. In certain embodiments, Form F is characterized as awhite powder. In certain embodiments, a sample of Form F comprisesparticles having a flake-like morphology. In certain embodiments, asample of Form F comprises particles with a D₉₀ of less than about 18μm.

Certain embodiments herein provide Form F of Compound A which issubstantially pure. Certain embodiments herein provide Form F ofCompound A which is substantially free of other solid forms comprisingCompound A including, e.g., Forms A, B, C, D, E, G and/or an amorphoussolid form comprising Compound A as provided herein. Certain embodimentsherein provide Form F as a mixture of solid forms comprising Compound A,including, e.g., a mixture comprising one or more of the following:Forms A, B, C, D, E, G and an amorphous solid form comprising Compound Aas provided herein.

4.1.7. Form G of Compound A

Certain embodiments herein provide the Form G crystal form of CompoundA. In certain embodiments, Form G of Compound A can be obtained fromvarious solvents, including, but not limited to, solvent systemscomprising ethyl acetate. For example, in certain embodiments, Form Gcan be obtained by crystallization from a solvent system comprisingethyl acetate, e.g., by a process comprising contacting a solid formcomprising Compound A with a solvent system comprising ethyl acetate,followed by isolation of Form G.

In certain embodiments, Form G of Compound A may be characterized byX-ray powder diffraction analysis. A representative XRPD pattern of FormG of Compound A is provided in FIG. 25. In certain embodiments, Form Gof Compound A is characterized by XRPD peaks located at one, two, three,four, five, six, seven, eight, nine, ten, eleven or twelve of thefollowing approximate positions: 7.9, 9.5, 11.7, 15.7, 16.8, 18.1, 19.7,21.8, 22.8, 25.1, 25.8, 26.7 degrees 2θ. In certain embodiments, Form Gof Compound A is characterized by an XRPD pattern which matches thepattern exhibited in FIG. 25. In certain embodiments, Form G of CompoundA is characterized by an XRPD pattern having 1, 2, 3, 4, 5, 6, 7, 8, 9,10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24 or 25 peaksmatching peaks in the representative Form G pattern provided herein.

In certain embodiments, Form G of Compound A may be characterized bythermal analysis. A representative DSC plot for Form G of Compound A isshown in FIG. 26. In certain embodiments, Form G is characterized by aDSC plot comprising an endothermic event with an onset temperature ofabout 109° C. A representative TGA plot for Form G of Compound A isshown in FIG. 27. In certain embodiments, Form G is characterized by aTGA plot comprising a mass loss of less than about 8%, e.g., about 3.8%,of the total mass of the sample upon heating from about 25° C. to about110° C. In certain embodiments, the TGA mass loss event comprises theloss of the solvent ethyl acetate, as indicated, e.g., by TG-IRanalysis. In certain embodiments, Form G of Compound A is solvated. Incertain embodiments, Form G is an ethyl acetate solvate. In certainembodiments, the crystal lattice of Form G comprises about three molarequivalents of ethyl acetate per mole of Compound A.

In certain embodiments, Form G of Compound A may be characterized bymoisture sorption analysis. A representative moisture sorption isothermplot is shown in FIG. 28. In certain embodiments, when the RH isincreased from about 0% to about 95% RH, Form G exhibits a mass changeof less than about 1%, e.g., about 0.4%, of the starting mass of thesample. In certain embodiments, mass gained upon adsorption is lost whenthe RH is decreased back to about 0% RH. In certain embodiments, Form Gis substantially nonhygroscopic. In certain embodiments, the XRPDpattern of Form G material is substantially unchanged following theadsorption/desorption analysis. In certain embodiments, Form G is stablewith respect to humidity.

In certain embodiments, Form G of Compound A may be characterized by itsstability profile. In certain embodiments, Form G material is stable,e.g., its XRPD pattern remains substantially unchanged, uponcompression. For example, in certain embodiments, Form F is stablefollowing compression at about 2000 psi pressure for about one minute.In certain embodiments, Form G converts to Form B upon exposure to asolvent system comprising, e.g., ethanol, acetone or mixtures thereof,for about two days at about 25° C.

In certain embodiments, Form G of Compound A may be characterized byparticle analysis. In certain embodiments, Form G is characterized as awhite powder. In certain embodiments, a sample of Form G comprisesparticles having a flake-like morphology. In certain embodiments, asample of Form G comprises particles with a D₉₀ of less than about 18μm.

Certain embodiments herein provide Form G of Compound A which issubstantially pure. Certain embodiments herein provide Form G ofCompound A which is substantially free of other solid forms comprisingCompound A including, e.g., Forms A, B, C, D, E, F and/or an amorphoussolid form comprising Compound A as provided herein. Certain embodimentsherein provide Form G as a mixture of solid forms comprising Compound A,including, e.g., a mixture comprising one or more of the following:Forms A, B, C, D, E, F and an amorphous solid form comprising Compound Aas provided herein.

4.2. Methods of Treatment

The invention encompasses methods of treating, preventing and managingdiseases or disorders ameliorated by the reduction of levels of TNF-α ina patient which comprise administering to a patient in need of suchtreatment, prevention or management a therapeutically orprophylactically effective amount of one or more solid forms comprisingCompound A, such as, e.g., Form A of Compound A, Form B of Compound A,Form C of Compound A, Form D of Compound A, Form E of Compound A, Form Fof Compound A, Form G of Compound A, or an amorphous form of Compound A,as provided herein.

Disorders ameliorated by the inhibition of TNF-α include, but are notlimited to: heart disease, such as congestive heart failure,cardiomyopathy, pulmonary edema, endotoxin-mediated septic shock, acuteviral myocarditis, cardiac allograft rejection, and myocardialinfarction; solid tumors, including but not limited to, sarcoma,carcinomas, fibrosarcoma, myxosarcoma, liposarcoma, chondrosarcoma,osteogenic sarcoma, chordoma, angiosarcoma, endotheliosarcoma,lymphangiosarcoma, lymphangioendotheliosarcoma, synovioma, mesothelioma,Ewing's tumor, leiomyosarcoma, rhabdomyosarcoma, colon carcinoma,pancreatic cancer, breast cancer, ovarian cancer, prostate cancer,squamous cell carcinoma, basal cell carcinoma, adenocarcinoma, sweatgland carcinoma, sebaceous gland carcinoma, papillary carcinoma,papillary adenocarcinomas, cystadenocarcinoma, medullary carcinoma,bronchogenic carcinoma, renal cell carcinoma, hepatoma, bile ductcarcinoma, choriocarcinoma, seminoma, embryonal carcinoma, Wilms' tumor,cervical cancer, testicular tumor, lung carcinoma, small cell lungcarcinoma, bladder carcinoma, epithelial carcinoma, glioma, astrocytoma,medulloblastoma, craniopharyngioma, ependymoma, Kaposi's sarcoma,pinealoma, hemangioblastoma, acoustic neuroma, oligodendroglioma,menangioma, melanoma, neuroblastoma, and retinoblastoma; and blood-bornetumors including but not limited to, acute lymphoblastic leukemia “ALL”,acute lymphoblastic B-cell leukemia, acute lymphoblastic T-cellleukemia, acute myeloblastic leukemia “AML”, acute promyelocyticleukemia “APL”, acute monoblastic leukemia, acute erythroleukemicleukemia, acute megakaryoblastic leukemia, acute myelomonocyticleukemia, acute nonlymphocyctic leukemia, acute undifferentiatedleukemia, chronic myelocytic leukemia “CML”, chronic lymphocyticleukemia “CLL”, hairy cell leukemia, multiple myeloma and acute andchronic leukemias, for example, lymphoblastic, myelogenous, lymphocytic,and myelocytic leukemias.

Specific methods of the invention further comprise the administration ofan additional therapeutic agent (i.e., a therapeutic agent other thanCompound A). Examples of additional therapeutic agents include, but arenot limited to, anti-cancer drugs such as, but are not limited to:alkylating agents, nitrogen mustards, ethylenimines, methylmelamines,alkyl sulfonates, nitrosoureas, triazenes, folic acid analogs,pyrimidine analogs, purine analogs, vinca alkaloids,epipodophyllotoxins, antibiotics, topoisomerase inhibitors andanti-cancer vaccines.

Specific additional therapeutic agents include, but are not limited to:acivicin; aclarubicin; acodazole hydrochloride; acronine; adozelesin;aldesleukin; altretamine; ambomycin; ametantrone acetate;aminoglutethimide; amsacrine; anastrozole; anthramycin; asparaginase;asperlin; azacitidine; azetepa; azotomycin; batimastat; benzodepa;bicalutamide; bisantrene hydrochloride; bisnafide dimesylate; bizelesin;bleomycin sulfate; brequinar sodium; bropirimine; busulfan;cactinomycin; calusterone; caracemide; carbetimer; carboplatin;carmustine; carubicin hydrochloride; carzelesin; cedefingol;chlorambucil; cirolemycin; cisplatin; cladribine; crisnatol mesylate;cyclophosphamide; cytarabine; dacarbazine; dactinomycin; daunorubicinhydrochloride; decitabine; dexormaplatin; dezaguanine; dezaguaninemesylate; diaziquone; docetaxel; doxorubicin; doxorubicin hydrochloride;droloxifene; droloxifene citrate; dromostanolone propionate; duazomycin;edatrexate; eflornithine hydrochloride; elsamitrucin; enloplatin;enpromate; epipropidine; epirubicin hydrochloride; erbulozole;esorubicin hydrochloride; estramustine; estramustine phosphate sodium;etanidazole; etoposide; etoposide phosphate; etoprine; fadrozolehydrochloride; fazarabine; fenretinide; floxuridine; fludarabinephosphate; fluorouracil; fluorocitabine; fosquidone; fostriecin sodium;gemcitabine; gemcitabine hydrochloride; hydroxyurea; idarubicinhydrochloride; ifosfamide; ilmofosine; interleukin II (includingrecombinant interleukin II, or rIL2), interferon alfa-2a; interferonalfa-2b; interferon alfa-n1; interferon alfa-n3; interferon beta-I a;interferon gamma-I b; iproplatin; irinotecan hydrochloride; lanreotideacetate; letrozole; leuprolide acetate; liarozolc hydrochloride;lometrexol sodium; lomustine; losoxantrone hydrochloride; masoprocol;maytansine; mechlorethamine hydrochloride; megestrol acetate;melengestrol acetate; melphalan; menogaril; mercaptopurine;methotrexate; methotrexate sodium; metoprine; meturedepa; mitindomide;mitocarcin; mitocromin; mitogillin; mitomalcin; mitomycin; mitosper;mitotane; mitoxantrone hydrochloride; mycophenolic acid; nocodazole;nogalamycin; ormaplatin; oxisuran; paclitaxel; pegaspargase; peliomycin;pentamustine; peplomycin sulfate; perfosfamide; pipobroman; piposulfan;piroxantrone hydrochloride; plicamycin; plomestane; porfimer sodium;porfiromycin; prednimustine; procarbazine hydrochloride; puromycin;puromycin hydrochloride; pyrazofurin; riboprine; rogletimide; safingol;safingol hydrochloride; semustine; simtrazene; sparfosate sodium;sparsomycin; spirogermanium hydrochloride; spiromustine; spiroplatin;streptonigrin; streptozocin; sulofenur; talisomycin; tecogalan sodium;tegafur; teloxantrone hydrochloride; temoporfin; teniposide; teroxirone;testolactone; thiamiprine; thioguanine; thiotepa; tiazofurin;tirapazamine; toremifene citrate; trestolone acetate; triciribinephosphate; trimetrexate; trimetrexate glucuronate; triptorelin;tubulozole hydrochloride; uracil mustard; uredepa; vapreotide;verteporfin; vinblastine sulfate; vincristine sulfate; vindesine;vindesine sulfate; vinepidine sulfate; vinglycinate sulfate;vinleurosine sulfate; vinorelbine tartrate; vinrosidine sulfate;vinzolidine sulfate; vorozole; zeniplatin; zinostatin; zorubicinhydrochloride. Other anti-cancer drugs include, but are not limited to:20-epi-1,25 dihydroxyvitamin D3; 5-ethynyluracil; abiraterone;aclarubicin; acylfulvene; adecypenol; adozelesin; aldesleukin; ALL-TKantagonists; altretamine; ambamustine; amidox; amifostine;aminolevulinic acid; amrubicin; amsacrine; anagrelide; anastrozole;andrographolide; angiogenesis inhibitors; antagonist D; antagonist G;antarelix; anti-dorsalizing morphogenetic protein-1; antiandrogen,prostatic carcinoma; antiestrogen; antineoplaston; antisenseoligonucleotides; aphidicolin glycinate; apoptosis gene modulators;apoptosis regulators; apurinic acid; ara-CDP-DL-PTBA; argininedeaminase; asulacrine; atamestane; atrimustine; axinastatin 1;axinastatin 2; axinastatin 3; azasetron; azatoxin; azatyrosine; baccatinIII derivatives; balanol; batimastat; BCR/ABL antagonists;benzochlorins; benzoylstaurosporine; beta lactam derivatives;beta-alethine; betaclamycin B; betulinic acid; bFGF inhibitor;bicalutamide; bisantrene; bisaziridinylspermine; bisnafide; bistrateneA; bizelesin; breflate; bropirimine; budotitane; buthionine sulfoximine;calcipotriol; calphostin C; camptothecin derivatives; canarypox IL-2;capecitabine; carboxamide-amino-triazole; carboxyamidotriazole; CaRestM3; CARN 700; cartilage derived inhibitor; carzelesin; casein kinaseinhibitors (ICOS); castanospermine; cecropin B; cetrorelix; chlorins;chloroquinoxaline sulfonamide; cicaprost; cis-porphyrin; cladribine;clomifene analogues; clotrimazole; collismycin A; collismycin B;combretastatin A4; combretastatin analogue; conagenin; crambescidin 816;crisnatol; cryptophycin 8; cryptophycin A derivatives; curacin A;cyclopentanthraquinones; cycloplatam; cypemycin; cytarabine ocfosfate;cytolytic factor; cytostatin; dacliximab; decitabine; dehydrodidemnin B;deslorelin; dexamethasone; dexifosfamide; dexrazoxane; dexverapamil;diaziquone; didemnin B; didox; diethylnorspermine;dihydro-5-azacytidine; dihydrotaxol, 9-; dioxamycin; diphenylspiromustine; docetaxel; docosanol; dolasetron; doxifluridine;droloxifene; dronabinol; duocarmycin SA; ebselen; ecomustine;edelfosine; edrecolomab; eflornithine; elemene; emitefur; epirubicin;epristeride; estramustine analogue; estrogen agonists; estrogenantagonists; etanidazole; etoposide phosphate; exemestane; fadrozole;fazarabine; fenretinide; filgrastim; finasteride; flavopiridol;flezelastine; fluasterone; fludarabine; fluorodaunorunicinhydrochloride; forfenimex; formestane; fostriecin; fotemustine;gadolinium texaphyrin; gallium nitrate; galocitabine; ganirelix;gelatinase inhibitors; gemcitabine; glutathione inhibitors; hepsulfam;heregulin; hexamethylene bisacetamide; hypericin; ibandronic acid;idarubicin; idoxifene; idramantone; ilmofosine; ilomastat;imidazoacridones; imiquimod; immunostimulant peptides; insulin-likegrowth factor-1 receptor inhibitor; interferon agonists; interferons;interleukins; iobenguane; iododoxorubicin; ipomeanol, 4-; iroplact;irsogladine; isobengazole; isohomohalicondrin B; itasetron;jasplakinolide; kahalalide F; lamellarin-N triacetate; lanreotide;leinamycin; lenograstim; lentinan sulfate; leptolstatin; letrozole;leukemia inhibiting factor; leukocyte alpha interferon;leuprolide+estrogen+progesterone; leuprorelin; levamisole; liarozole;linear polyamine analogue; lipophilic disaccharide peptide; lipophilicplatinum compounds; lissoclinamide 7; lobaplatin; lombricine;lometrexol; lonidamine; losoxantrone; lovastatin; loxoribine;lurtotecan; lutetium texaphyrin; lysofylline; lytic peptides;maitansine; mannostatin A; marimastat; masoprocol; maspin; matrilysininhibitors; matrix metalloproteinase inhibitors; menogaril; merbarone;meterelin; methioninase; metoclopramide; MIF inhibitor; mifepristone;miltefosine; mirimostim; mismatched double stranded RNA; mitoguazone;mitolactol; mitomycin analogues; mitonafide; mitotoxin fibroblast growthfactor-saporin; mitoxantrone; mofarotene; molgramostim; monoclonalantibody, human chorionic gonadotrophin; monophosphoryl lipidA+myobacterium cell wall sk; mopidamol; multiple drug resistance geneinhibitor; multiple tumor suppressor 1-based therapy; mustard anticanceragent; mycaperoxide B; mycobacterial cell wall extract; myriaporone;N-acetyldinaline; N-substituted benzamides; nafarelin; nagrestip;naloxone+pentazocine; napavin; naphterpin; nartograstim; nedaplatin;nemorubicin; neridronic acid; neutral endopeptidase; nilutamide;nisamycin; nitric oxide modulators; nitroxide antioxidant; nitrullyn;O6-benzylguanine; octreotide; okicenone; oligonucleotides; onapristone;ondansetron; ondansetron; oracin; oral cytokine inducer; ormaplatin;osaterone; oxaliplatin; oxaunomycin; paclitaxel; paclitaxel analogues;paclitaxel derivatives; palauamine; palmitoylrhizoxin; pamidronic acid;panaxytriol; panomifene; parabactin; pazelliptine; pegaspargase;peldesine; pentosan polysulfate sodium; pentostatin; pentrozole;perflubron; perfosfamide; perillyl alcohol; phenazinomycin;phenylacetate; phosphatase inhibitors; picibanil; pilocarpinehydrochloride; pirarubicin; piritrexim; placetin A; placetin B;plasminogen activator inhibitor; platinum complex; platinum compounds;platinum-triamine complex; porfimer sodium; porfiromycin; prednisone;propyl bis-acridone; prostaglandin J2; proteasome inhibitors; proteinA-based immune modulator; protein kinase C inhibitor; protein kinase Cinhibitors, microalgal; protein tyrosine phosphatase inhibitors; purinenucleoside phosphorylase inhibitors; purpurins; pyrazoloacridine;pyridoxylated hemoglobin polyoxyethylene conjugate; raf antagonists;raltitrexed; ramosetron; ras farnesyl protein transferase inhibitors;ras inhibitors; ras-GAP inhibitor; retelliptine demethylated; rhenium Re186 etidronate; rhizoxin; ribozymes; RII retinamide; rogletimide;rohitukine; romurtide; roquinimex; rubiginone B1; ruboxyl; safingol;saintopin; SarCNU; sarcophytol A; sargramostim; Sdi 1 mimetics;semustine; senescence derived inhibitor 1; sense oligonucleotides;signal transduction inhibitors; signal transduction modulators; singlechain antigen binding protein; sizofiran; sobuzoxane; sodiumborocaptate; sodium phenylacetate; solverol; somatomedin bindingprotein; sonermin; sparfosic acid; spicamycin D; spiromustine;splenopentin; spongistatin 1; squalamine; stem cell inhibitor; stem-celldivision inhibitors; stipiamide; stromelysin inhibitors; sulfinosine;superactive vasoactive intestinal peptide antagonist; suradista;suramin; swainsonine; synthetic glycosaminoglycans; tallimustine;tamoxifen methiodide; tauromustine; tazarotene; tecogalan sodium;tegafur; tellurapyrylium; telomerase inhibitors; temoporfin;temozolomide; teniposide; tetrachlorodecaoxide; tetrazomine;thaliblastine; thiocoraline; thrombopoietin; thrombopoietin mimetic;thymalfasin; thymopoietin receptor agonist; thymotrinan; thyroidstimulating hormone; tin ethyl etiopurpurin; tirapazaminc; titanocenebichloride; topsentin; toremifene; totipotent stem cell factor;translation inhibitors; tretinoin; triacetyluridine; triciribine;trimetrexate; triptorelin; tropisetron; turosteride; tyrosine kinaseinhibitors; tyrphostins; UBC inhibitors; ubenimex; urogenitalsinus-derived growth inhibitory factor; urokinase receptor antagonists;vapreotide; variolin B; vector system, erythrocyte gene therapy;velaresol; veramine; verdins; verteporfin; vinorelbine; vinxaltine;vitaxin; vorozole; zanoterone; zeniplatin; zilascorb; and zinostatinstimalamer.

Embodiments herein further encompass a method of treating or preventingdiseases or disorders ameliorated by the inhibition of PDE4 in a patientwhich comprise administering to a patient in need of such treatment orprevention one or more solid forms comprising Compound A. Disordersameliorated by the inhibition of PDE4 include, but are not limited to,asthma, inflammation, chronic or acute obstructive pulmonary disease,chronic or acute pulmonary inflammatory disease, inflammatory boweldisease, Crohn's Disease, Behcet's Disease, colitis, ulcerative colitisand arthritis or inflammation due to reperfusion. In a preferredembodiment, the disease or disorder to be treated or prevented ischronic obstructive pulmonary disease.

Specific methods of the invention can comprise the administration of anadditional therapeutic agent such as, but not limited to,anti-inflammatory drugs, antihistamines and decongestants. Examples ofsuch additional therapeutic agents include, but are not limited to:antihistamines including, but not limited to, ethanolamines,ethylenediamines, piperazines, and phenothiazines; antinflammatorydrugs; NSAIDS, including, but not limited to, aspirin, salicylates,acetominophen, indomethacin, sulindac, etodolac, fenamates, tolmetin,ketorolac, diclofenac, ibuprofen, naproxen, fenoprofen, ketoprofen,flurbiprofen, oxaprozin, piroxicam, meloxicam, pyrazolon derivatives;and steroids including, but not limited to, cortical steroids andadrenocortical steroids.

Specific methods of the invention avoid or reduce drug-drug interactionsand other adverse effects associated with agents used in the treatmentof such disorders, including racemic substituted phenylethylsulfones.Without being limited by any theory, certain solid forms comprisingCompound A may further provide an overall improved therapeuticeffectiveness, or therapeutic index, over racemic2-[1-(3-Ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dione,including solid forms thereof.

As stated above, certain solid forms comprising Compound A may be usedin the treatment or prevention of a wide range of diseases andconditions. The magnitude of a prophylactic or therapeutic dose of aparticular active ingredient of the invention in the acute or chronicmanagement of a disease or condition may vary with the nature andseverity of the disease or condition and the route by which the activeingredient is administered. The dose, and perhaps the dose frequency,will also vary according to the age, body weight, and response of theindividual patient. Suitable dosing regimens can be readily selected bythose skilled in the art with due consideration of such factors. Ingeneral, the recommended daily dose range for the conditions describedherein lie within the range of from about 1 mg to about 1,000 mg perday, given as a single once-a-day dose preferably as divided dosesthroughout a day. More specifically, the daily dose is administeredtwice daily in equally divided doses. Specifically, a daily dose rangemay be from about 5 mg to about 500 mg per day, more specifically,between about 10 mg and about 200 mg per day. Specifically, the dailydose may be administered in 5 mg, 10 mg, 15 mg, 20 mg, 25 mg, 50 mg, or100 mg dosage forms. In managing the patient, the therapy should beinitiated at a lower dose, perhaps about 1 mg to about 25 mg, andincreased if necessary up to about 200 mg to about 1,000 mg per day aseither a single dose or divided doses, depending on the patient's globalresponse. Alternatively, the daily dose is from 0.01 mg/kg to 100 mg/kg.

It may be necessary to use dosages of the active ingredient outside theranges disclosed herein in some cases, as will be apparent to those ofordinary skill in the art. Furthermore, it is noted that the clinicianor treating physician will know how and when to interrupt, adjust, orterminate therapy in conjunction with individual patient response.

The phrases “therapeutically effective amount”, “prophylacticallyeffective amount” and “therapeutically or prophylactically effectiveamount,” as used herein encompass the above described dosage amounts anddose frequency schedules. Different therapeutically effective amountsmay be applicable for different diseases and conditions, as will bereadily known by those of ordinary skill in the art. Similarly, amountssufficient to treat or prevent such disorders, but insufficient tocause, or sufficient to reduce, adverse effects associated with racemic2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetylaminoisoindoline-1,3-dioneare also encompassed by the above described dosage amounts and dosefrequency schedules.

4.3. Pharmaceutical Compositions

Pharmaceutical compositions and single unit dosage forms comprising oneor more solid forms comprising Compound A are provided herein. Alsoprovided herein are methods for preparing pharmaceutical compositionsand single unit dosage forms comprising one or more solid formscomprising Compound A. For example, in certain embodiments, individualdosage forms comprising a solid form provided herein or prepared usingsolid form provided herein may be suitable for oral, mucosal (includingrectal, nasal, or vaginal), parenteral (including subcutaneous,intramuscular, bolus injection, intraarterial, or intravenous),sublingual, transdermal, buccal, or topical administration.

In certain embodiments, pharmaceutical compositions and dosage formsprovided herein comprise one or more solid forms comprising Compound A.Certain embodiments herein provide pharmaceutical compositions anddosage forms comprising a solid form comprising Compound A, such as,e.g., Forms A, B, C, D, E, F, G or an amorphous solid form comprisingCompound A as provided herein, wherein the solid form comprisingCompound A substantially pure. Certain embodiments herein providepharmaceutical compositions and dosage forms comprising a solid formcomprising Compound A, such as, e.g., Forms A, B, C, D, E, F, G or anamorphous solid form comprising Compound A as provided herein, which issubstantially free of other solid forms comprising Compound A including,e.g., Forms A, B, C, D, E, F, G and/or an amorphous solid formcomprising Compound A as provided herein. Certain embodiments hereinprovide pharmaceutical compositions and dosage forms comprising amixture of solid forms comprising Compound A, including, e.g., a mixturecomprising one or more of the following: Forms A, B, C, D, E, F and anamorphous solid form comprising Compound A as provided herein.Pharmaceutical compositions and dosage forms provided herein typicallyalso comprise one or more pharmaceutically acceptable excipient, diluentor carrier.

A particular pharmaceutical composition encompassed by this embodimentcomprises one or more solid forms comprising Compound A and at least oneadditional therapeutic agent. Examples of additional therapeutic agentsinclude, but are not limited to: anti-cancer drugs and anti-inflammationtherapies including, but not limited to, those provided herein.

Single unit dosage forms of the invention are suitable for oral, mucosal(e.g., nasal, sublingual, vaginal, buccal, or rectal), parenteral (e.g.,subcutaneous, intravenous, bolus injection, intramuscular, orintraarterial), or transdermal administration to a patient. Examples ofdosage forms include, but are not limited to: tablets; caplets;capsules, such as soft elastic gelatin capsules; cachets; troches;lozenges; dispersions; suppositories; ointments; cataplasms (poultices);pastes; powders; dressings; creams; plasters; solutions; patches;aerosols (e.g., nasal sprays or inhalers); gels; liquid dosage formssuitable for oral or mucosal administration to a patient, includingsuspensions (e.g., aqueous or non-aqueous liquid suspensions,oil-in-water emulsions, or a water-in-oil liquid emulsions), solutions,and elixirs; liquid dosage forms suitable for parenteral administrationto a patient; and sterile solids (e.g., crystalline or amorphous solids)that can be reconstituted to provide liquid dosage forms suitable forparenteral administration to a patient.

The composition, shape, and type of dosage forms of the invention willtypically vary depending on their use. For example, a dosage form usedin the acute treatment of inflammation or a related disorder may containlarger amounts of one or more of the active ingredients it comprisesthan a dosage form used in the chronic treatment of the same disease.Similarly, a parenteral dosage form may contain smaller amounts of oneor more of the active ingredients it comprises than an oral dosage formused to treat the same disease or disorder. These and other ways inwhich specific dosage forms encompassed by this invention will vary fromone another will be readily apparent to those skilled in the art. See,e.g., Remington's Pharmaceutical Sciences, 18th ed., Mack Publishing,Easton Pa. (1990).

Typical pharmaceutical compositions and dosage forms comprise one ormore excipients. Suitable excipients are well known to those skilled inthe art of pharmacy, and non-limiting examples of suitable excipientsare provided herein. Whether a particular excipient is suitable forincorporation into a pharmaceutical composition or dosage form dependson a variety of factors well known in the art including, but not limitedto, the way in which the dosage form will be administered to a patient.For example, oral dosage forms such as tablets may contain excipientsnot suited for use in parenteral dosage forms. The suitability of aparticular excipient may also depend on the specific active ingredientsin the dosage form.

Lactose-free compositions of the invention can comprise excipients thatare well known in the art and are listed, for example, in the U.S.Pharmocopia (USP) SP (XXI)/NF (XVI). In general, lactose-freecompositions comprise an active ingredient, a binder/filler, and alubricant in pharmaceutically compatible and pharmaceutically acceptableamounts. Preferred lactose-free dosage forms comprise an activeingredient, microcrystalline cellulose, pre-gelatinized starch, andmagnesium stearate.

This invention further encompasses anhydrous pharmaceutical compositionsand dosage forms comprising active ingredients, since water canfacilitate the degradation of some compounds. For example, the additionof water (e.g., 5%) is widely accepted in the pharmaceutical arts as ameans of simulating long-term storage in order to determinecharacteristics such as shelf-life or the stability of formulations overtime. See, e.g., Jens T. Carstensen, Drug Stability: Principles &Practice, 2d. Ed., Marcel Dekker, NY, N.Y., 1995, pp. 379-80. In effect,water and heat accelerate the decomposition of some compounds. Thus, theeffect of water on a formulation can be of great significance sincemoisture and/or humidity are commonly encountered during manufacture,handling, packaging, storage, shipment, and use of formulations.

Anhydrous pharmaceutical compositions and dosage forms of the inventioncan be prepared using anhydrous or low moisture containing ingredientsand low moisture or low humidity conditions. Pharmaceutical compositionsand dosage forms that comprise lactose and at least one activeingredient that comprises a primary or secondary amine are preferablyanhydrous if substantial contact with moisture and/or humidity duringmanufacturing, packaging, and/or storage is expected.

An anhydrous pharmaceutical composition should be prepared and storedsuch that its anhydrous nature is maintained. Accordingly, anhydrouscompositions are preferably packaged using materials known to preventexposure to water such that they can be included in suitable formularykits. Examples of suitable packaging include, but are not limited to,hermetically sealed foils, plastics, unit dose containers (e.g., vials),blister packs, and strip packs.

The invention further encompasses pharmaceutical compositions and dosageforms that comprise one or more compounds that reduce the rate by whichan active ingredient will decompose. Such compounds, which are referredto herein as “stabilizers,” include, but are not limited to,antioxidants such as ascorbic acid, pH buffers, or salt buffers.

Like the amounts and types of excipients, the amounts and specific typesof active ingredients in a dosage form may differ depending on factorssuch as, but not limited to, the route by which it is to be administeredto patients. However, typical dosage forms provided herein lie withinthe range of from about 1 mg to about 1,000 mg per day, given as asingle once-a-day dose in the morning but preferably as divided dosesthroughout the day. More specifically, the daily dose is administeredtwice daily in equally divided doses. Specifically, a daily dose rangemay be from about 5 mg to about 500 mg per day, more specifically,between about 10 mg and about 200 mg per day. In managing the patient,the therapy may be initiated at a lower dose, perhaps about 1 mg toabout 25 mg, and increased if necessary up to about 200 mg to about1,000 mg per day as either a single dose or divided doses, depending onthe patient's global response.

4.3.1. Oral Dosage Forms

Pharmaceutical compositions of the invention that are suitable for oraladministration can be presented as discrete dosage forms, such as, butare not limited to, tablets (e.g., chewable tablets), caplets, capsules,and liquids (e.g., flavored syrups). Such dosage forms containpredetermined amounts of active ingredients, and may be prepared bymethods of pharmacy well known to those skilled in the art. Seegenerally Remington's Pharmaceutical Sciences, 18th ed., MackPublishing, Easton Pa. (1990).

Typical oral dosage forms of the invention are prepared by combining theactive ingredient(s) in an intimate admixture with at least oneexcipient according to conventional pharmaceutical compoundingtechniques. Excipients can take a wide variety of forms depending on theform of preparation desired for administration. For example, excipientssuitable for use in oral liquid or aerosol dosage forms include, but arenot limited to, water, glycols, oils, alcohols, flavoring agents,preservatives, and coloring agents. Examples of excipients suitable foruse in solid oral dosage forms (e.g., powders, tablets, capsules, andcaplets) include, but are not limited to, starches, sugars,micro-crystalline cellulose, diluents, granulating agents, lubricants,binders, and disintegrating agents.

Because of their ease of administration, tablets and capsules representthe most advantageous oral dosage unit forms, in which case solidexcipients are employed. If desired, tablets can be coated by standardaqueous or nonaqueous techniques. Such dosage forms can be prepared byany of the methods of pharmacy. In general, pharmaceutical compositionsand dosage forms are prepared by uniformly and intimately admixing theactive ingredients with liquid carriers, finely divided solid carriers,or both, and then shaping the product into the desired presentation ifnecessary.

For example, a tablet can be prepared by compression or molding.Compressed tablets can be prepared by compressing in a suitable machinethe active ingredients in a free-flowing form such as powder orgranules, optionally mixed with an excipient. Molded tablets can be madeby molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent.

Examples of excipients that can be used in oral dosage forms of theinvention include, but are not limited to, binders, fillers,disintegrants, and lubricants. Binders suitable for use inpharmaceutical compositions and dosage forms include, but are notlimited to, corn starch, potato starch, or other starches, gelatin,natural and synthetic gums such as acacia, sodium alginate, alginicacid, other alginates, powdered tragacanth, guar gum, cellulose and itsderivatives (e.g., ethyl cellulose, cellulose acetate, carboxymethylcellulose calcium, sodium carboxymethyl cellulose), polyvinylpyrrolidone, methyl cellulose, pre-gelatinized starch, hydroxypropylmethyl cellulose, (e.g., Nos. 2208, 2906, 2910), microcrystallinecellulose, and mixtures thereof.

Examples of fillers suitable for use in the pharmaceutical compositionsand dosage forms disclosed herein include, but are not limited to, talc,calcium carbonate (e.g., granules or powder), microcrystallinecellulose, powdered cellulose, dextrates, kaolin, mannitol, silicicacid, sorbitol, starch, pre-gelatinized starch, and mixtures thereof.The binder or filler in pharmaceutical compositions of the invention istypically present in from about 50 to about 99 weight percent of thepharmaceutical composition or dosage form.

Suitable forms of microcrystalline cellulose include, but are notlimited to, the materials sold as AVICEL-PH-101™, AVICEL-PH-103™, AVICELRC-581™, AVICEL-PH-105™ (available from FMC Corporation, AmericanViscose Division, Avicel Sales, Marcus Hook, Pa.), and mixtures thereof.A specific binder is a mixture of microcrystalline cellulose and sodiumcarboxymethyl cellulose sold as AVICEL RC-581™. Suitable anhydrous orlow moisture excipients or additives include AVICEL-PH-103™ and Starch1500 LM™.

Disintegrants are used in the compositions of the invention to providetablets that disintegrate when exposed to an aqueous environment.Tablets that contain too much disintegrant may disintegrate in storage,while those that contain too little may not disintegrate at a desiredrate or under the desired conditions. Thus, a sufficient amount ofdisintegrant that is neither too much nor too little to detrimentallyalter the release of the active ingredients should be used to form solidoral dosage forms of the invention. The amount of disintegrant usedvaries based upon the type of formulation, and is readily discernible tothose of ordinary skill in the art. Typical pharmaceutical compositionscomprise from about 0.5 to about 15 weight percent of disintegrant,specifically from about 1 to about 5 weight percent of disintegrant.

Disintegrants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, agar-agar,alginic acid, calcium carbonate, microcrystalline cellulose,croscarmellose sodium, crospovidone, polacrilin potassium, sodium starchglycolate, potato or tapioca starch, pre-gelatinized starch, otherstarches, clays, other algins, other celluloses, gums, and mixturesthereof.

Lubricants that can be used in pharmaceutical compositions and dosageforms of the invention include, but are not limited to, calciumstearate, magnesium stearate, mineral oil, light mineral oil, glycerin,sorbitol, mannitol, polyethylene glycol, other glycols, stearic acid,sodium lauryl sulfate, talc, hydrogenated vegetable oil (e.g., peanutoil, cottonseed oil, sunflower oil, sesame oil, olive oil, corn oil, andsoybean oil), zinc stearate, ethyl oleate, ethyl laureate, agar, andmixtures thereof. Additional lubricants include, for example, a syloidsilica gel (AEROSIL 200™, manufactured by W.R. Grace Co. of Baltimore,Md.), a coagulated aerosol of synthetic silica (marketed by Degussa Co.of Plano, Tex.), CAB-O-SIL™ (a pyrogenic silicon dioxide product sold byCabot Co. of Boston, Mass.), and mixtures thereof. If used at all,lubricants are typically used in an amount of less than about one weightpercent of the pharmaceutical compositions or dosage forms into whichthey are incorporated.

4.3.2. Delayed Release Dosage Forms

Solid forms comprising Compound A as provided herein can be administeredby controlled release means or by delivery devices that are well knownto those of ordinary skill in the art. Examples include, but are notlimited to, those described in U.S. Pat. Nos. 3,845,770; 3,916,899;3,536,809; 3,598,123; and 4,008,719, 5,674,533, 5,059,595, 5,591,767,5,120,548, 5,073,543, 5,639,476, 5,354,556, and 5,733,566, each of whichis incorporated herein by reference. Such dosage forms can be used toprovide slow or controlled-release of one or more active ingredientsusing, for example, hydropropylmethyl cellulose, other polymer matrices,gels, permeable membranes, osmotic systems, multilayer coatings,microparticles, liposomes, microspheres, or a combination thereof toprovide the desired release profile in varying proportions. Suitablecontrolled-release formulations known to those of ordinary skill in theart, including those described herein, can be readily selected for usewith the active ingredients of the invention. The invention thusencompasses single unit dosage forms suitable for oral administrationsuch as, but not limited to, tablets, capsules, gelcaps, and capletsthat are adapted for controlled-release.

All controlled-release pharmaceutical products have a common goal ofimproving drug therapy over that achieved by their non-controlledcounterparts. Ideally, the use of an optimally designedcontrolled-release preparation in medical treatment is characterized bya minimum of drug substance being employed to cure or control thecondition in a minimum amount of time. Advantages of controlled-releaseformulations include extended activity of the drug, reduced dosagefrequency, and increased patient compliance. In addition,controlled-release formulations can be used to affect the time of onsetof action or other characteristics, such as blood levels of the drug,and can thus affect the occurrence of side (e.g., adverse) effects.

Most controlled-release formulations are designed to initially releasean amount of drug (active ingredient) that promptly produces the desiredtherapeutic effect, and gradually and continually release of otheramounts of drug to maintain this level of therapeutic or prophylacticeffect over an extended period of time. In order to maintain thisconstant level of drug in the body, the drug must be released from thedosage form at a rate that will replace the amount of drug beingmetabolized and excreted from the body. Controlled-release of an activeingredient can be stimulated by various conditions including, but notlimited to, pH, temperature, enzymes, water, or other physiologicalconditions or compounds.

4.3.3. Parenteral Dosage Forms

Parenteral dosage forms can be administered to patients by variousroutes including, but not limited to, subcutaneous, intravenous(including bolus injection), intramuscular, and intraarterial. Becausetheir administration typically bypasses patients' natural defensesagainst contaminants, parenteral dosage forms are preferably sterile orcapable of being sterilized prior to administration to a patient.Examples of parenteral dosage forms include, but are not limited to,solutions ready for injection, dry products ready to be dissolved orsuspended in a pharmaceutically acceptable vehicle for injection,suspensions ready for injection, and emulsions.

Suitable vehicles that can be used to provide parenteral dosage forms ofthe invention are well known to those skilled in the art. Examplesinclude, but are not limited to: Water for Injection USP; aqueousvehicles such as, but not limited to, Sodium Chloride Injection,Ringer's Injection, Dextrose Injection, Dextrose and Sodium ChlorideInjection, and Lactated Ringer's Injection; water-miscible vehicles suchas, but not limited to, ethyl alcohol, polyethylene glycol, andpolypropylene glycol; and non-aqueous vehicles such as, but not limitedto, corn oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate,isopropyl myristate, and benzyl benzoate.

Compounds that increase the solubility of one or more of the activeingredients disclosed herein can also be incorporated into theparenteral dosage forms of the invention.

4.3.4. Transdermal, Topical, and Mucosal Dosage Forms

Transdermal, topical, and mucosal dosage forms of the invention include,but are not limited to, ophthalmic solutions, sprays, aerosols, creams,lotions, ointments, gels, solutions, emulsions, suspensions, or otherforms known to one of skill in the art. See, e.g., Remington'sPharmaceutical Sciences, 16th and 18th eds., Mack Publishing, Easton Pa.(1980 & 1990); and Introduction to Pharmaceutical Dosage Forms, 4th ed.,Lea & Febiger, Philadelphia (1985). Dosage forms suitable for treatingmucosal tissues within the oral cavity can be formulated as mouthwashesor as oral gels. Further, transdermal dosage forms include “reservoirtype” or “matrix type” patches, which can be applied to the skin andworn for a specific period of time to permit the penetration of adesired amount of active ingredients.

Suitable excipients (e.g., carriers and diluents) and other materialsthat can be used to provide transdermal, topical, and mucosal dosageforms encompassed by this invention are well known to those skilled inthe pharmaceutical arts, and depend on the particular tissue to which agiven pharmaceutical composition or dosage form will be applied. Withthat fact in mind, typical excipients include, but are not limited to,water, acetone, ethanol, ethylene glycol, propylene glycol,butane-1,3-diol, isopropyl myristate, isopropyl palmitate, mineral oil,and mixtures thereof to form lotions, tinctures, creams, emulsions, gelsor ointments, which are non-toxic and pharmaceutically acceptable.Moisturizers or humectants can also be added to pharmaceuticalcompositions and dosage forms if desired. Examples of such additionalingredients are well known in the art. See, e.g., Remington'sPharmaceutical Sciences, 16th and 18th eds., Mack Publishing, Easton Pa.(1980 & 1990).

Depending on the specific tissue to be treated, additional componentsmay be used prior to, in conjunction with, or subsequent to treatmentwith active ingredients of the invention. For example, penetrationenhancers can be used to assist in delivering the active ingredients tothe tissue. Suitable penetration enhancers include, but are not limitedto: acetone; various alcohols such as ethanol, olcyl, andtetrahydrofuryl; alkyl sulfoxides such as dimethyl sulfoxide; dimethylacetamide; dimethyl formamide; polyethylene glycol; pyrrolidones such aspolyvinylpyrrolidone; Kollidon grades (Povidone, Polyvidone); urea; andvarious water-soluble or insoluble sugar esters such as Tween 80™(polysorbate 80) and Span 60™ (sorbitan monostearate).

The pH of a pharmaceutical composition or dosage form, or of the tissueto which the pharmaceutical composition or dosage form is applied, mayalso be adjusted to improve delivery of one or more active ingredients.Similarly, the polarity of a solvent carrier, its ionic strength, ortonicity can be adjusted to improve delivery. Compounds such asstearates can also be added to pharmaceutical compositions or dosageforms to advantageously alter the hydrophilicity or lipophilicity of oneor more active ingredients so as to improve delivery. In this regard,stearates can serve as a lipid vehicle for the formulation, as anemulsifying agent or surfactant, and as a delivery-enhancing orpenetration-enhancing agent. Different solid forms comprising the activeingredients can be used to further adjust the properties of theresulting composition.

4.3.5. Kits

This invention encompasses kits which, when used by the medicalpractitioner, can simplify the administration of appropriate amounts ofactive ingredients to a patient.

A typical kit of the invention comprises a unit dosage form of compoundA, or a pharmaceutically acceptable solid form or prodrug thereof, and aunit dosage form of a second active ingredient. Examples of secondactive ingredients include, but are not limited to, those listed herein.

Kits of the invention can further comprise devices that are used toadminister the active ingredient(s). Examples of such devices include,but are not limited to, syringes, drip bags, patches, and inhalers.

Kits of the invention can further comprise pharmaceutically acceptablevehicles that can be used to administer one or more active ingredients.For example, if an active ingredient is provided in a solid form thatmust be reconstituted for parenteral administration, the kit cancomprise a sealed container of a suitable vehicle in which the activeingredient can be dissolved to form a particulate-free sterile solutionthat is suitable for parenteral administration. Examples ofpharmaceutically acceptable vehicles include, but are not limited to:Water for Injection USP; aqueous vehicles such as, but not limited to,Sodium Chloride Injection, Ringer's Injection, Dextrose Injection,Dextrose and Sodium Chloride Injection, and Lactated Ringer's Injection;water-miscible vehicles such as, but not limited to, ethyl alcohol,polyethylene glycol, and polypropylene glycol; and non-aqueous vehiclessuch as, but not limited to, corn oil, cottonseed oil, peanut oil,sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate.

5. EXAMPLES

The present application incorporates by reference the entirety of U.S.Pat. No. 6,962,940 (issued Nov. 8, 2005), including the Examplesprovided therein.

5.1. Example 1 SYNTHESIS OF2-[1-(3-ETHOXY-4-METHOXYPHENYL)-2-METHYLSULFONYLETHYL]-4-ACETYLAMINOISOINDOLINE-1,3-DIONE

A stirred solution of1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethylamine (1.0 g, 3.7mmol) and 3-acetamidophthalic anhydride (751 mg, 3.66 mmol) in aceticacid (20 mL) was heated at reflux for 15 h. The solvent was removed invacuo to yield an oil. Chromatography of the resulting oil yielded theproduct as a yellow solid (1.0 g, 59% yield): mp, 144° C.; ¹H NMR(CDCl₃) δ: 1.47 (t, J=7.0 Hz, 3H, CH₃), 2.26 (s, 3H, CH₃), 2.88 (s, 3H,CH₃), 3.75 (dd, J=4.4, 14.3 Hz, 1H, CH), 3.85 (s, 3H, CH3), 4.11 (q, J=7Hz, 2H, CH₂), 5.87 (dd, J=4.3, 10.5 Hz, 1H, NCH), 6.82-6.86 (m, 1H, Ar),7.09-7.11 (m, 2H, Ar), 7.47 (d, J=7 Hz, 1H, Ar), 7.64 (t, J=8 Hz, 1H,Ar), 8.74 (d, J=8 Hz, 1H, Ar), 9.49 (br s, 1H, NH); ¹³C NMR (CDCl₃) δ:14.61, 24.85, 41.54, 48.44, 54.34, 55.85, 64.43, 111.37, 112.34, 115.04,118.11, 120.21, 124.85, 129.17, 130.96, 136.01, 137.52, 148.54, 149.65,167.38, 169.09, 169.40; Anal Calc'd. for C₂₂H₂₄NO₇S: C, 57.38; H, 5.25;N, 6.08. Found: C, 57.31; H, 5.34; N, 5.83.

5.2. Example 2 SYNTHESIS OF(+)2-[1-(3-ETHOXY-4-METHOXYPHENYL)-2-METHYLSULFONYLETHYL]-4-ACETYLAMINOISOINDOLINE-1,3-DIONEPreparation of 3-aminopthalic acid

10% Pd/C (2.5 g), 3-nitrophthalic acid (75.0 g, 355 mmol) and ethanol(1.5 L) were charged to a 2.5 L Parr hydrogenator under a nitrogenatmosphere. Hydrogen was charged to the reaction vessel for up to 55psi. The mixture was shaken for 13 hours, maintaining hydrogen pressurebetween 50 and 55 psi. Hydrogen was released and the mixture was purgedwith nitrogen 3 times. The suspension was filtered through a celite bedand rinsed with methanol. The filtrate was concentrated in vacuo. Theresulting solid was reslurried in ether and isolated by vacuumfiltration. The solid was dried in vacuo to a constant weight, affording54 g (84% yield) of 3-aminopthalic acid as a yellow product. ¹H-NMR(DMSO-d6) δ: 3.17 (s, 2H), 6.67 (d, 1H), 6.82 (d, 1H), 7.17 (t, 1H),8-10 (br, s, 2H); ¹³C-NMR (DMSO-d6) δ: 112.00, 115.32, 118.20, 131.28,135.86, 148.82, 169.15, 170.09.

Preparation of 3-acetamidophthalic anhydride

A 1 L 3-necked round bottom flask was equipped with a mechanicalstirrer, thermometer, and condenser and charged with 3-aminophthalicacid (108 g, 596 mmol) and acetic anhydride (550 mL). The reactionmixture was heated to reflux for 3 hours and cooled to about 25° C. andfurther to 0-5° C. for another 1 hour. The crystalline solid wascollected by vacuum filtration and washed with ether. The solid productwas dried in vacuo at ambient temperature to a constant weight, giving75 g (61% yield) of 3-acetamidopthalic anhydride as a white product.¹H-NMR (CDCl₃) δ: 2.21 (s, 3H), 7.76 (d, 1H), 7.94 (t, 1H), 8.42 (d,1H), 9.84 (s, 1H).

Resolution of2-(3-ethoxy-4-methoxyphenyl-1-(methylsulphonyl)-eth-2-ylamine

A 3 L 3-necked round bottom flask was equipped with a mechanicalstirrer, thermometer, and condenser and charged with2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-ylamine (137.0 g,500 mmol), N-acetyl-L-leucine (52 g, 300 mmol), and methanol (1.0 L).The stirred slurry was heated to reflux for 1 hour. The stirred mixturewas allowed to cool to ambient temperature and stirring was continuedfor another 3 hours at ambient temperature. The slurry was filtered andwashed with methanol (250 L). The solid was air-dried and then dried invacuo at ambient temperature to a constant weight, giving 109.5 g (98%yield) of the crude product (85.8% ee). The crude solid (55.0 g) andmethanol (440 mL) were brought to reflux for 1 hour, cooled to roomtemperature and stirred for an additional 3 hours at ambienttemperature. The slurry was filtered and the filter cake was washed withmethanol (200 mL). The solid was air-dried and then dried in vacuo at30° C. to a constant weight, yielding 49.6 g (90% recovery) of(S)-2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-ylamine-N-acetyl-L-leucinesalt (98.4% ee). Chiral HPLC (1/99 EtOH/20 mM KH₂PO₄ @ pH 7.0, UltronChiral ES-OVS from Agilent Technologies, 150 mm×4.6 mm, 0.5 mL/min., @240 nm): 18.4 min (S-isomer, 99.2%), 25.5 min (R-isomer, 0.8%).

Preparation of Compound A

A 500 mL 3-necked round bottom flask was equipped with a mechanicalstirrer, thermometer, and condenser. The reaction vessel was chargedwith (S)-2-(3-ethoxy-4-methoxyphenyl)-1-(methylsulphonyl)-eth-2-yl amineN-acetyl-L-leucine salt (25 g, 56 mmol, 98% ee), 3-acetamidophthalicanhydride (12.1 g, 58.8 mmol), and glacial acetic acid (250 mL). Themixture was refluxed over night and then cooled to <50° C. The solventwas removed in vacuo, and the residue was dissolved in ethyl acetate.The resulting solution was washed with water (250 mL×2), saturatedaqueous NaHCO₃ (250 mL×2), brine (250 mL×2), and dried over sodiumsulphate. The solvent was evaporated in vacuo, and the residuerecrystallized from a binary solvent containing ethanol (150 mL) andacetone (75 mL). The solid was isolated by vacuum filtration and washedwith ethanol (100 mL×2). The product was dried in vacuo at 60° C. to aconstant weight, affording 19.4 g (75% yield) ofS-{2-[1-(3-ethoxy-4-methoxyphenyl)-2-methylsulfonylethyl]-4-acetamidoisoindoline-1,3-dione}with 98% ee. Chiral HPLC (15/85 EtOH/20 mM KH₂PO₄@ pH 5, Ultron ChiralES-OVS from Agilent Technology, 150 mm×4.6 mm, 0.4 mL/min, @ 240 nm):25.4 min (S-isomer, 98.7%), 29.5 min (R-isomer, 1.2%). (CDCl₃) δ: 1.47(t, 3H), 2.26 (s, 3H), 2.87 (s, 3H), 3.68-3.75 (dd, 1H), 3.85 (s, 3H),4.07-4.15 (q, 2H), 4.51-4.61 (dd, 1H), 5.84-5.90 (dd, 1H), 6.82-8.77 (m,6H), 9.46 (s, 1H); ¹³C-NMR (DMSO-d6) δ: 14.66, 24.92, 41.61, 48.53,54.46, 55.91, 64.51, 111.44, 112.40, 115.10, 118.20, 120.28, 124.94,129.22, 131.02, 136.09, 137.60, 148.62, 149.74, 167.46, 169.14, 169.48.

A reaction scheme illustrating a preparation of the (+) enantiomer ofCompound A is provided as FIG. 29.

5.3. Example 3 TNF-α Inhibition

Human Whole Blood LPS-Induced TNF-α Assay

The ability of compounds to inhibit LPS-induced TNF-α production byhuman whole blood was measured essentially as described below for theLPS-induced TNF-α assay in human PBMC, except that freshly drawn wholeblood was used instead of PBMC. (Muller et al., 1999, Bioorg. & Med.Chem. Lett., 9:1625-1630.) Human whole blood LPS-induced TNF-α IC₅₀=294nM for Compound A.

Mouse LPS-Induced Serum TNF-α Inhibition

Compounds were tested in this animal model according to previouslydescribed methods (Corral et al., 1996, Mol. Med., 2:506-515). MouseLPS-induced scrum TNF-α inhibition (ED₅₀, mg/kg, p.o.)=0.05 for CompoundA.

LPS-Induced TNF-α Production

Lipopolysaccharide (LPS) is an endotoxin produced by gram-negativebacteria such as E. coli which induces production of manypro-inflammatory cytokines, including TNF-α. In peripheral bloodmononuclear cells (PBMC), the TNF-α produced in response to LPS isderived from monocytes, which comprise approximately 5-20% of the totalPBMC. Compounds were tested for the ability to inhibit LPS-induced TNF-αproduction from human PBMC as previously described (Muller et al., 1996,J. Med. Chem., 39:3238). PBMC from normal donors were obtained by FicollHypaque (Pharmacia, Piscataway, N.J., USA) density centrifugation. Cellswere cultured in RPMI (Life Technologies, Grand Island, N.Y., USA)supplemented with 10% AB± human serum (Gemini Bio-products, Woodland,Calif., USA), 2 mM L-glutamine, 100 U/ml penicillin, and 100 μg/mlstreptomycin (Life Technologies).

PBMC (2×10⁵ cells) were plated in 96-well flat-bottom Costar tissueculture plates (Corning, N.Y., USA) in triplicate. Cells were stimulatedwith LPS (Sigma, St. Louis, Mo., USA) at 100 ng/ml in the absence orpresence of compounds. Compounds (Celgene Corp., Warren, N.J., USA) weredissolved in DMSO (Sigma) and further dilutions were done in culturemedium immediately before use. The final DMSO concentration in allsamples was 0.25%. Compounds were added to cells one hour before LPSstimulation. Cells were incubated for 18-20 hours at 37° C. in 5% CO₂and supernatants were then collected, diluted with culture medium andassayed for TNF-α levels by ELISA (Endogen, Boston, Mass., USA).LPS-induced TNF-α IC₅₀=77 nM for Compound A.

IL-1β-Induced TNF-α Production

During the course of inflammatory diseases, TNF-α production is oftenstimulated by the cytokine 1L-1β, rather than by bacterially derivedLPS. Compounds were tested for the ability to inhibit IL-1β-inducedTNF-α production from human PBMC as described above for LPS-inducedTNF-α production, except that the PBMC were isolated from sourceleukocyte units (Sera-Tec Biologicals, North Brunswick, N.J., USA) bycentrifugation on Ficoll-Paque Plus (Amersham Pharmacia, Piscataway,N.J., USA), plated in 96-well tissue culture plates at 3×10⁵ cells/wellin RPMI-1640 medium (BioWhittaker, Walkersville, Md., USA) containing10% heat-inactivated fetal bovine serum (Hyclone), 2 mM L-glutamine, 100U/ml penicillin, and 100 mg/ml streptomycin (complete medium),pretreated with compounds at 10, 2, 0.4, 0.08, 0.016, 0.0032, 0.00064,and 0 μM in duplicate at a final DMSO concentration of 0.1% at 37° C. ina humidified incubator at 5% CO₂ for one hour, then stimulated with 50ng/ml recombinant human IL-1β (Endogen) for 18 hours. IL-β-induced TNF-αIC₅₀=83 nM for Compound A.

5.4. Example 4 PDE Selectivity

PDE1, 2, 3, 5, and 6 Enzyme Assays

The specificity of compounds for PDE4 was assessed by testing at asingle concentration (10 μM) against bovine PDE1, human PDE2, PDE3, andPDE5 from human platelets (Hidaka and Asano, 1976, Biochem. Biophys.Acta, 429:485, and Nicholsen et al., 1991, Trends Pharmaco. Sci.,12:19), and PDE6 from bovine retinal rod outer segments (Baehr et al.,1979, J. Biol. Chem., 254:11669, and Gillespie et al. 1989, Mol. Pharm.,36:773). Results are listed in Table 1.

PDE7 Enzyme Assay

PDE7 is a cAMP-selective PDE expressed mainly in T cells and in skeletalmuscle. T cell-derived cytokines such as IL-2 and IFN-γ are potentiallyregulatable via PDE7 inhibition. PDE7 was purified from Hut78 human Tcells by anion exchange chromatography as previously described (Bloomand Beavo, 1996, Proc. Natl. Acad. Sci. USA, 93:14188-14192). Compoundswere tested against the PDE7 preparation in the presence of 10 nM cAMPas described for PDE4 in Table 1.

5.5. Example 5 PDE4 Inhibition

PDE4 (U937 Cell-Derived) Enzyme Assay

PDE4 enzyme was purified from U937 human monocytic cells by gelfiltration chromatography as previously described (Muller et al., 1998,Bioorg. & Med. Chem. Lett. 8:2669-2674). Phosphodiesterase reactionswere carried out in 50 mM Tris HCl pH 7.5, 5 mM MgCl₂, 1 μM cAMP, 10 nM[³H]-cAMP for 30 min at 30° C., terminated by boiling, treated with 1mg/ml snake venom, and separated using AG-1XS ion exchange resin(BioRad) as described (Muller et al., 1998, Bioorg. & Med. Chem. Lett.8:2669-2674). Reactions consumed less than 15% of available substrate.Results are listed in Table 1.

TABLE 1 PDE Specificity Racemic Compound Compound A Compound B* PDEInhibition PDE4 IC₅₀ (from U937 81.8 73.5 611 cells) (nM) PDE1 (% inhibat  9% 23% 27% 10 μM) PDE2(% inhib at 19%  6% 10% 10 μM) PDE3 (% inhibat 21% 20% 31% 10 μM) PDE5 (% inhib at  3%  3% −9% μM) PDE6 (% inhib atND −6% 10% 10 μM) PDE7 IC₅₀ (nM) 22110 20500 ND PDE Specificity Ratiosfrom above data (*fold) PDE4/PDE1 >2700 >500 >50PDE4/PDE2 >800 >10000 >260 PDE4/PDE3 >670 >1200 >45PDE4/PDE5 >12000 >30000 >39000 PDE4/PDE6 ND >40000 >250 PDE7 IC₅₀/PDE4IC₅₀ 270 279 ND *Compound B is the ( ) enantiomer of Compound A.

5.6. Example 6 Human T Cell Assays

SEB-Induced IL-2 and IFN-γ Production

Staphylococcal Enterotoxin B (SEB) is a superantigen derived fromgram-positive bacteria Staphylococcus aureus. SEB provides a convenientphysiological stimulus specific for T cells expressing particular T cellreceptor Vβ chains. Human PBMC (consisting of approximately 50% T cells)were isolated from source leukocyte units as described above and platedin 96-well tissue culture plates at 3×10⁵ cells/well in complete medium,pretreated with compounds at 10, 2, 0.4, 0.08, 0.016, 0.0032, 0.00064,and 0 μM in duplicate at a final DMSO concentration of 0.1% at 37° C. ina humidified incubator at 5% CO₂ for 1 hour, then stimulated with 100ng/ml SEB (Sigma Chemical Co., St. Louis, Mo., USA) for 18 hours. IL-2and IFN-γ levels were measured by ELISA (R&D Systems, Minneapolis,Minn., USA). IL-2 IC₅₀=291 nM for Compound A. IFN-γ IC₅₀=46 nM forCompound A.

5.7. Example 7 Camp Elevation Assays

PGE₂-Induced cAMP Elevation

Prostaglandin E₂ (PGE₂) binds to prostanoid receptors on monocytes, Tcells and other leukocytes and consequently elevates intracellular cAMPlevels, resulting in inhibition of cellular responses. The combinationof PGE₂ and a PDE4 inhibitor synergistically elevates cAMP levels inthese cell types, and the elevation of cAMP in PBMC caused by PDE4inhibitors in the presence of PGE₂ is proportional to the inhibitoryactivity of that PDE4 inhibitor. Intracellular cAMP was measured inhuman PBMC as follows. PBMC were isolated as described above and platedin 96-well plates at 1×10⁶ cells per well in RPMI-1640. The cells werepre-treated with compounds at 100, 10, 1, 0.1, 0.01, and 0 μM in a finalconcentration of 2% DMSO in duplicate at 37° C. in a humidifiedincubator at 5% CO₂ for one hour. The cells were then stimulated withPGE₂ (10 μM) (Sigma) for 1 h. The cells were lysed with HCl, 0.1 N finalconcentration to inhibit phosphodiesterase activity and the plates werefrozen at −20° C. The cAMP produced was measured using cAMP (low pH)Immunoassay kit (R&D Systems). PBMC cAMP EC₅₀ for racemate is 3.09 μM.PBMC cAMP EC₅₀ for Compound A is 1.58 μM.

Elevation of cAMP in human neutrophils was measured as follows. PBMCwere removed from source leukocytes (Sera-Tec Biologicals) bycentrifugation on Ficoll-Paque Plus (Amersham Pharmacia). The resultingerythrocyte/polymorphonuclear cell (PMN) pellet was resuspended inHank's Balanced Salt Solution (BioWhittaker) and mixed with an equalvolume of 3% Dextran T-500 (Amersham Pharmacia) in 0.9% saline.Erythrocytes were allowed to sediment for 20 minutes, and the PMN wereremoved and centrifuged at 120 rpm for 8 minutes at 4° C. The remainingerythrocytes were lysed in cold 0.2% saline for 30 seconds, and thecells restored to isotonicity by the addition of an equal volume of 1.6%saline. The PMN were centrifuged at 1200 rpm for 8 minutes at 4° C.,then resuspended in RPMI-1640 and assayed for cAMP elevation asdescribed for PBMC above. PMN were found to be approximately 74%CD18/CD11b⁺, 71% CD16⁺CD9⁺ neutrophils by flow cytometzy on aFACSCalibur (Becton Dickinson, San Jose, Calif., USA). Results are shownin Table 2.

fMLF-Induced LTB4 Production

N-formyl-methionine-leucine-phenylalanine (fMLF) is a bacteriallyderived peptide that activates neutrophils to rapidly degranulate,migrate, adhere to endothelial cells, and release leukotriene LTB4, aproduct of arachidonic acid metabolism and itself a neutrophilchemoattractant. Compounds were tested for the ability to blockfMLF-induced neutrophil LTB4 production as previously described(Hatzelmann and Schudt, 2001, J. Pharm. Exp. Ther., 297:267-279), withthe following modifications. Neutrophils were isolated as describedabove and resuspended in phosphate-buffered saline without calcium ormagnesium (BioWhittaker) containing 10 mM HEPES pH 7.2 and plated in96-well tissue culture plates at a concentration of 1.7×10⁶ cells/well.Cells were treated with 50 μM thimerosal (Sigma)/1 mM CaCl₂/1 mM MgCl₂for 15 minutes at 37° C. 5% CO₂, then treated with compounds at 1000,200, 40, 8, 1.6, 0.32, 0.064, and 0 nM in a final DMSO concentration of0.01% in duplicate for 10 minutes. Neutrophils were stimulated with 1 μMfMLF for 30 minutes, then lysed by the addition of methanol (20% finalconcentration) and frozen in a dry ice/isopropanol bath for 10 minutes.Lysates were stored at −70° C. until the LTB4 content was measured bycompetitive LTB4 ELISA (R&D Systems). Results are shown in Table 2.

Zymosan-Induced IL-8 Production

Zymosan A, or the heat-killed yeast Saccharomyces cerevisiae, binds tothe adhesion molecule Mac-1 on the neutrophil surface and triggersphagocytosis, cell activation and IL-8 production. Zymosan-induced IL-8production was measured as previously described (Au et al., 1998, Brit.J. Pharm., 123:1260-1266) with the following modifications. Humanneutrophils were purified as described above, plated in 96-well tissueculture plates at 3×10⁵ cells/well in complete medium, treated withcompounds at 10, 2, 0.4, 0.08, 0.016, 0.0032, 0.00064, and 0 μM induplicate in a final DMSO concentration of 0.1% for 1 hour at 37° C. 5%CO₂. Neutrophils were then stimulated with unopsonized, boiled Zymosan A(Sigma) at 2.5×10⁵ particles/well for 18 hours. Supernatants wereharvested and tested for IL-8 by ELISA (R&D Systems). Results are shownin Table 2.

fMLF-Induced CDI8/CD11b Expression

CD18/CD11b (Mac-1) expression on neutrophils was measured as previouslydescribed (Derian et al., 1995, J. Immunol., 154:308-3 17) with thefollowing modifications. Neutrophils were isolated as described above,then resuspended in complete medium at 1×10⁶ cells/ml, pretreated withcompounds at 10, 1, 0.1, 0.01, and 0 μM in duplicate at a final DMSOconcentration of 0.1% for 10 minutes at 37° C. 5% CO₂. Cells were thenstimulated with 30 nM fMLF for 30 minutes and then chilled to 4° C.Cells were treated with rabbit IgG (Jackson ImmunoResearch Labs, WestGrove, Pa., USA) (10 μg/1×10⁶ cells) to block Fc receptors, stained withCD18-FITC and CD11b-PE (Becton Dickinson), and analyzed by flowcytometry on a FACSCalibur. CD18/CD11b expression (mean fluorescence) inthe absence of stimulation was subtracted from all samples to obtaininhibition curves and calculate IC₅₀ values. Results are shown in Table2.

fMLF-Induced Adhesion to HUVEC

Human umbilical vein endothelial cells (HUVEC) were used as a substratefor neutrophil adhesion as previously described (Derian et al., 1995, J.Immunol., 154:308-317) with the following modifications. HUVEC cellswere obtained from Anthrogenesis (Cedar Knolls, N.J., USA), andneutrophils were not treated with cytochalasin B. Cells were treatedwith compounds at 10, 1, 0.1, 0.01, 0.001, and 0 μM in a final DMSOconcentration of 0.1% in duplicate for 10 minutes, stimulated with 500nM fMLF for 30 minutes, and washed twice with PBS before measuringfluorescence on an FLX800 plate reader (Bio-Tek Instruments, Winooski,Vt., USA). Results are shown in Table 2.

TABLE 2 Assay results Human Neutrophil Assays Racemic (all values in nM)Compound Compound A PGE₂-induced cAMP EC₅₀ 12589 4570 fMLF-induced LTB4IC₅₀ 20.1 2.48 Zymosan-induced IL-8 IC₅₀ ND 94 fMLF-induced CD18expression IC₅₀ ND 390 fMLF-induced CD11b expression IC₅₀ ND 74fMLF-induced adhesion to HUVEC IC₅₀ ND 150

5.8. Example 8 Aqueous Solubility

Equilibrium solubilities were measured in pH 7.4 aqueous buffer. The pH7.4 buffer was prepared by adjusting the pH of a 0.07 M NaH₂PO₄ solutionto 7.4 with 10 N NaOH. The ionic strength of the solution was 0.15. Atleast 1 mg of powder was combined with 1 ml of buffer to make >1 mg/mlmixture. These samples were shaken for >2 hours and left to standovernight at room temperature. The samples were then filtered through a0.45-1 μm Nylon syringe filter that was first saturated with the sample.The filtrate was sampled twice, consecutively. The filtrate was assayedby HPLC against standards prepared in 50% methanol. Compound A has3.5-fold greater aqueous solubility than the racemic mixture. Measuredsolubility Compound A=0.012 mg/mL; racemic mixture=0.0034 mg/mL.

5.9. Example 9 LPS-Induced Lung Neutrophilia Ferret Model

The conscious ferret model has been used to investigateanti-inflammatory, emetic and behavioral effects of PDE4 inhibitors whenadministered by the oral (p.o.) route. From these experiments, atherapeutic index (TI) for each PDE4 inhibitor may be determined. The TIhas been calculated by dividing the threshold dose for causing emeticepisodes and behavioral changes by the anti-inflammatory dose (dose thatcauses 50% inhibition of the LPS-induced neutrophilia).

Animal Husbandry

Male ferrets (Mustela Pulorius Euro, weighing 1-2 kg). Ferrets weresupplied either by Bury Green Farm or Misay Consultancy. Followingtransport, the animals were allowed to acclimatize in the holding roomsfor a period of not less than seven days. The diet comprised SDS diet Cpelleted food given ad lib with Whiskers™ cat food given three times perweek. Water was pasteurized animal grade drinking water and was changeddaily.

Dosing with PDE4 Inhibitor

PDE4 inhibitors were administered orally (p.o.), at doses initially of1-10 g/kg, but subsequently up to 30 mg/kg in order to establish whetherthe TI was 10 or higher, and/or at lower doses to establish the minimumdose to cause 50% inhibition of neutrophilia. Ferrets were fastedovernight but allowed free access to water. The animals were orallydosed with vehicle or PDE4 inhibitor using a 15 cm dosing needle thatwas passed down the back of the throat into the oesophagus. Afterdosing, the animals were returned to holding cages fitted with Perspexdoors to allow observation, and given free access to water. Afterdosing, the animals were constantly observed and any emesis orbehavioral changes were recorded. The animals were allowed access tofood 60 to 90 minutes after p.o. dosing.

Exposure to LPS

Thirty minutes after p.o. dosing with compound or vehicle control, theferrets were placed into sealed Perspex containers and exposed to anaerosol of LPS (100 μg/ml) for 10 minutes. Aerosols of LPS weregenerated by a nebulizer (DeVilbiss, USA) and this was directed into thePerspex exposure chamber. Following a 10 minute exposure period, theanimals were returned to the holding cages and allowed free access towater, and at a later stage, food. Observation continued for a period ofat least 2.5 hours post p.o. dosing and emetic episodes and behavioralchanges were recorded.

Bronchoalveolar Lavage

Six hours after LPS exposure the animals were killed by overdose ofsodium pentobarbitone administered intraperitoneally. The trachea wasthen cannulated with polypropylene tubing and the lungs lavaged twicewith 20 ml heparinized (10 units/ml) phosphate buffered saline (PBS).

Blood Sampling/Tissue Removal

A terminal blood sample (10 ml) was removed by trans-thoracic cardiacpuncture. The blood was spun at 2,500 rpm for 15 minutes and the plasmawas removed and stored at −20° C. The brain also removed and frozen at−20° C. for analysis of compound content.

Cell Counts

The bronchoalveolar lavage (BAL) samples were centrifuged at 1,500 rpmfor 5 minutes. The supernatant was removed and the resulting cell pelletre-suspended in 1 ml PBS. A cell smear of the re-suspended fluid wasprepared and stained with Leishmans stain to allow differential cellcounting. A total cell count was made using the remaining re-suspendedsample. From this, the total number of neutrophils in the BAL wasdetermined.

Parameters Measured

1. % Inhibition of LPS-induced pulmonary neutrophilia.

2. Emetic episodes—the number of vomits and retches were counted.

3. Behavioral changes—the following behavioral effects were noted:salivation, panting, mouth clawing, flattened posture, ataxia, archedback and backward walking. Any behavioral changes were semi-quantifiedby applying a severity rating (mild, moderate or severe).

4. The TI was calculated as the highest dose found to not cause emeticepisodes divided by the lowest dose found to inhibit pulmonaryneutrophilia by 50% or more.

The effect of Compound A on LPS-induced neutrophilia in the lungs ofconscious ferrets is demonstrated in FIG. 30.

Emesis and Behavioral Changes

Following p.o. dosing of the PDE4, the ferrets were observed for atleast two hours and emetic episodes (vomits and retches) and behavioralchanges were recorded.

No emetic episodes (retching or vomiting) were observed in the ferretspre-treated p.o. with the relevant vehicle (acetone/cremophor/distilledwater). In a small proportion of the control-treated animals (7/22),mild behavioral changes (lip licking and backward walking) were seen.

Compound A (0.1-3 mg/kg, p.o.), caused no emetic episodes (retching andvomiting). Some behavioral changes (flattened posture, lip licking andbackward walking) were observed and classified as mild. At 10 mg/kg in2/6 ferrets, some retching but no frank emesis was observed along withsalivation and behavioral changes (scored as mild or moderate). At thehighest dose tested (30 mg/kg) moderate to marked emesis was observed in3/4 animals along with pronounced behavioral changes. These data aresummarized in Table 3.

TABLE 3 Conscious ferret: Emetic episodes and behavioral changesfollowing oral administration of Compound A Treatment/dose MouthFlattened Backward (mg/kg) Vomits Retches Salivation Panting clawingposture Ataxia Lip licking walking Vehicle None None None None None NoneNone Mild Mild (acetone/ (6/22) (7/22) cremophor/ dist. H₂O) Compound ANone None None None None Mild None Mild Mild (0.1 mg/kg) (2/5) (4/5)(3/5) Compound A None None None None None Mild None Mild Mild (0.3mg/kg) (2/6) (3/6) (4/6) Compound A None None None None None Mild NoneMild Mild (1.0 mg/kg) (2/6) (6/6) (4/6) Compound A None None None NoneMild Marked None Mild Moderate (3.0 mg/kg) (1/8) (7/8) (2/8) (5/8)Compound A None Mild Mild (1/6) None Mild Marked None Moderate Marked(10 mg/kg) (2/6) (1/6) (6/6) (5/6) (6/6) Compound A Moderate MarkedModerate Mild Marked Marked Mild Moderate Mild (30 mg/kg) (3/4) (3/4)(3/4) (1/4) (4/4) (4/4) (3/4) (4/4) (2/4)

Animals were observed for up to three hours following dosing. Numbers inparentheses refer to the number of animals that responded. The numbersof animals in each group range from 4 to 22.

Therapeutic Index Calculation

From these experiments, a therapeutic index (TI) was determined for eachcompound by dividing the threshold dose for inducing emetic episodes bythe ED₅₀ value for inhibiting the pulmonary neutrophilia. The TIcalculation is summarized in Table 4. Compound A had a TI of 12, causingno emetic episodes at an anti-inflammatory dose of 1 mg/kg.

TABLE 4 Summary of the effective doses (ED₅₀) for inhibition ofLPS-induced pulmonary neutrophilia and induction of emesis and thetherapeutic index derived from these values Inhibition of LPS-inducedneutrophilia Threshold emetic Therapeutic Compound (ED₅₀ mg/kg) dose(mg/kg) index Compound A 0.8 10 12

5.10. Example 10 Biological Activity of Compound A in Patients withSevere Plaque-Type Psoriasis

Compound A is a novel oral agent that downregulates pro-inflammatorycytokine production in human cellular models. Compound A has been shownto decrease TNF-α, IL-12 and IFN-γ production as well as elevateproduction of IL-10. Psoriasis is strongly associated with dysregulationof cytokines and chemokines allowing for potential therapies withimmunomodulatory compounds. This Phase 2, open-label, single arm, pilotstudy was designed to assess the biological activity of Compound A inpatients with severe plaque-type psoriasis. Additional assessments forclinical outcomes were performed to evaluate the potential efficacy ofCompound A in treating severe plaque-type psoriasis.

Compound A was administered 20 mg orally daily for 29 days with anadditional 28-day observational follow-up period for patient safety.Skin punch biopsy specimens (6 mm) from target plaques were obtained atbaseline, Day 15 and Day 29. A nonlesional skin biopsy was also taken atbaseline. The primary pharmacodynamic endpoint was the percent changefrom baseline in epidermal thickness at Day 29. Epidermal skin thicknessmeasurements and immunohistochemical analysis were carried out by ablinded reviewer to evaluate CD11c, CD83, K16, ICAM-1, HLA-DR, andfillagrin. Biopsy specimens were analyzed by RT-PCR for: TNF-α,p40-IL12/IL23, IL-10, IFN-γ, IP10, IL-2, IL-8, iNOS, p19-IL23, K16, CD83, and hARP. PASI, PGA, and BSA measurements were performed to exploreclinical efficacy during the 29-day treatment phase of the study.Adverse event reporting, clinical laboratory evaluations, physicalexaminations, ECG and vital sign measurements assessed safety. A totalof 19 patients were enrolled: 15 patients had complete sets of evaluablebiopsies and 17 patients had complete efficacy assessments.

Assessment of the change in epidermal thickness was the primary endpointin this study. Nineteen patients were enrolled in the study, of which 15had complete sets of evaluable biopsies at baseline and Day 29.Seventeen of the 19 subjects had clinical efficacy parameters measuredat Baseline and Day 29. Eight (53.3%) of the patients with evaluablebiopsies at baseline and Day 29 demonstrated a 20% reduction inepidermal skin thickness. The mean reduction of epidermal thicknessamong all 15 subjects with evaluable biopsies at baseline and Day 29 was20.5% (p=0.015). FIG. 31 displays the change in epidermal thickness frombaseline to Day 29 among subjects with evaluable biopsies.

Key inflammatory markers including epidermal and dermal T-cells, CD83+and CD11c cells were evaluated in biopsy specimens. Results for 8patients who responded showed a decrease of epidermal and dermal T-cellsby 42.56% and 28.79% respectively in responders (≦20% epidermalthickness reduction). Mean reductions from baseline in epidermal anddermal CD83+ cells were 32.50% and 25.86% respectively in responders.CD11c cells were reduced by 40.16% in the epidermis and 18.50% in thedermis in responders. Table 5 lists reductions in key skin biopsyinflammatory markers in responders and nonresponders. In addition, onepatient with abnormal K16 at baseline had normal K16 at Day 29. Threepatients with abnormal ICAM-1 at baseline had normal ICAM-1 at Day 29.Two patients with abnormal HLA-DR had normal HLA-DR at Day 29 and threepatients with abnormal fillagrin at baseline had normal fillagrin at Day29.

TABLE 5 Percentage Reduction of Key Inflammatory Markers at Day 29 CellEpidermis Dermis T-cells Responder −42.56% −28.79% Nonresponder +8.74%−17.34% CD83+ Responder −32.50% −25.86% Nonresponder −16.31% +0.46%CD11c Responder −40.16% −18.50% Nonresponder −2.54% −21.19%

Biopsy specimens were evaluated for mRNA gene expression of keyinflammatory markers by RT-PCR including: TNFα, p40-IL12/IL23, IL-10,IFNγ, IP10, IL-2, IL-8, iNOS, p19-IL23, K16 and CD83. The mRNAexpression of iNOS was reduced 66.5% (p=0.025) in lesional skin after 29days of treatment with Compound A. Reductions and increases in mRNAexpression of other inflammatory markers showed overall trends ofimprovement. FIG. 32 graphically displays the change in iNOS expressionduring the study.

A total of 17 of the 19 subjects enrolled completed the 29-day treatmentphase and had complete clinical efficacy assessments. Fourteen (73.7%)of the 19 subjects enrolled demonstrated improvement in their PASI with3 (15.8%) of these patients showing a >50% reduction from baseline intheir total Psoriasis Area and Severity Index (PASI) score at Day 29.FIG. 33 displays the percentage change in PASI scores among evaluablepatients from baseline at Day 29. Additionally, 9 (52.9%) of the 17evaluable patients demonstrated improvement in the static Physician'sGlobal Assessment (sPGA) and 10 (58.8%) of the 17 evaluable patientsshowed a reduction from baseline in their psoriasis body surface area(BSA) after 29 days of treatment with Compound A. Safety was evaluatedduring treatment and follow-up phases through monitoring of adverseevents, ECGs, laboratory tests, physical exams and vital signs. Nodeaths were reported nor did any patient prematurely discontinue due toan adverse event. Most common treatment-related adverse events includedheadache (26.3%), and nausea (15.8%).

In this clinical study, Compound A 20 mg p.o. QD for 29 days was safe insubjects with severe plaquetype psoriasis. The primary endpoint wasreached with 8 (53.3%) of 15 subjects achieving a 20% reduction inepidermal thickness at Day 29. Reductions of key inflammatory markers inskin biopsies were noted including dermal and epidermal T-cells, CD83+and CD11c cells. RT-PCR analysis revealed a statistically significantreduction of 66.5% in iNOS mRNA in skin biopsies at Day 29. A positiveclinical efficacy signal was noted after 29 days of treatment withCompound A. 73.7% of enrolled patients demonstrated improvement in theirpsoriasis symptoms with 15.8% of these patients showing >50% reductionfrom baseline in their PASI score at Day 29. 47.4% of enrolled patientsshowed an improvement in their sPGA and 52.6% of enrolled patientsshowed a reduction from baseline in their psoriasis body surface area(BSA) at Day 29.

5.11. Example 11 A Phase 2 Study Demonstrating the Efficacy and Safetyof Compound A in Subjects with Moderate-to-Severe Psoriasis

This phase 2, multicenter, randomized, double-blind, placebo-controlled,parallel-group, dose-comparison study evaluated the efficacy and safetyof Compound A in subjects with moderate to severe plaque-type psoriasiswho were candidates for systemic therapy.

This study included a 12-week treatment phase followed by a 4-weekobservational follow-up phase. A total of 260 subjects were randomizedto receive Compound A 20 mg BID, Compound A 20 mg QD, or placebo for 12weeks. The primary endpoint for this study was the proportion ofsubjects treated with Compound A who achieved a 75% reduction inPsoriasis Area and Severity Index score (“PASI-75”) at week 12/lasttreatment in reference to the baseline visit. Last treatment is definedas the last PAST assessment completed during the 12-week treatmentphase.

At week 12/last treatment, a significantly higher proportion of subjectstreated with 20 mg BID (24%) achieved a PASI-75 compared with theplacebo group (10%; P=0.023). Of the subjects receiving 20 mg BID orplacebo, 57% versus 23% achieved PASI-50 at week 12/last treatment,respectively; whereas 14% versus 6% achieved PASI-90, respectively. Atweek 12/last treatment, subjects achieved a mean decrease of 52% versus17% in PASI from baseline in the 20 mg BID versus placebo groups,respectively. Subjects receiving Compound A continued to improve overtime, showing the greatest mean percent reduction in PASI score at week12. Overall, the adverse event profiles were similar across all threetreatment groups. The majority of adverse events reported were mild. Nostudy drug-related serious adverse events were reported in this study.No subjects in the 20 mg BID group experienced psoriasis flare duringthe observational follow-up period.

In this clinical study, Compound A was shown to be well tolerated andsafe in subjects with moderate to severe plaque-type psoriasis. Theproportions of subjects that achieved 50%, 75%, and 90% improvement inPASI demonstrate the clinical activity of Compound A after 12 weeks oftreatment.

5.12. Example 12 Solid Form Screening Studies

5.12.1. Experimental Methodology

Solubility Studies.

A weighed sample of Compound A (about 100 mg) was treated with about 2mL of the test solvent. The solvents used were either reagent or HPLCgrade. The resulting mixture was agitated for at least 24 hours at about25° C. When all of the solids were dissolved by visual inspection, theestimated solubilities were calculated. The solubilities were estimatedfrom these experiments based on the total volume of solvent used to givea solution. The actual solubilities may be greater than those calculateddue to the use of large amount of solvent or to a slow rate ofdissolution. If dissolution did not occur during the experiment, thesolubility was measured gravimetrically. A known volume of filtrate wasevaporated to dryness and the weight of the residue was measured.

Solution Evaporation Studies.

Solution evaporation was performed for solvents in which the solubilityof Compound A was more than about 50 mg/mL, such as acetone,acetonitrile, methylene chloride and tetrahydrofuran. Solid samples wereobtained by slowly evaporating the solvents at about 25° C. or about 50°C. in an open vial under nitrogen.

Equilibration Studies.

Equilibration experiments were carried out by adding an excess ofCompound A to about 2 mL of a test solvent. The resulting mixture wasagitated for at least 24 hours at about 25° C. or about 50° C. Uponreaching equilibrium, the saturated solution was removed and allowed toevaporate slowly in an open vial under nitrogen at about 25° C. or about50° C., respectively. The slurry resulting from the equilibration wasfiltered and dried in the air.

Cooling Crystallization Studies.

Cooling crystallization studies were performed. The solid was dissolvedin a solvent at an elevated temperature, about 65° C., and allowed tocool to about 25° C. Samples that did not crystallize at about 25° C.were placed in a refrigerator (about 0-5° C.). Solids were isolated bydecantation and allowed to dry in the air.

Solvent/Anti-Solvent Precipitation Studies.

Precipitations were carried out by solvent/anti-solvent combinations.The solid was dissolved in a solvent in which Compound A had arelatively high solubility, and then a selected solvent in whichCompound A had a relatively low solubility (i.e., an anti-solvent) wasadded to the solution. A precipitate formed immediately in somesolvent/anti-solvent systems. If the precipitation did not occurimmediately, the resulting mixture was allowed to cool in a refrigerator(about 0-5° C.) until a precipitate formed. The precipitate was thenisolated by decantation and allowed to dry in the air.

Interconversion Studies.

Interconversion experiments were performed by making slurries of a solidform in a saturated solvent. The slurries were agitated for at least 2days at about 25° C. The saturated solution was removed by filtrationand the solid was dried in the air.

Compression Studies.

Compression tests were performed by pressing the sample under 2000 psiforce for at least one minute with Carver Mini C presser. The sample wasthen analyzed by XRPD.

Hygroscopicity Studies.

The hygroscopicity of various solid forms was studied using a SurfaceMeasurement Systems DVS instrument. Typically a sample size of betweenabout 10-50 mg was loaded into the DVS instrument sample pan and thesample was analyzed on a DVS automated sorption analyzer at about 25° C.The relative humidity was increased in increments of about 10% fromabout 0% to about 95% RH. The relative humidity was then decreased in asimilar manner to accomplish a full adsorption/desorption cycle. Themass was recorded at periodic intervals throughout the experiment.

5.12.2. Characterization Methodology

Samples generated as described in the solid form screen were typicallyanalyzed by X-Ray Powder Diffraction (XRPD). XRPD was conducted on aThermo ARL X'TRA™ X-ray powder diffractometer using Cu Kα radiation at1.54 Å. The instrument was equipped with a fine focus X-ray tube. Thevoltage and amperage of X-ray generator were set at 45 kV and 40 mA,respectively. The divergence slices were set at 4 mm and 2 mm and themeasuring slices were set at 0.5 mm and 0.2 mm. The diffracted radiationwas detected by a peltier-cooled Si(Li) solid-state detector. Typically,a theta-two theta continuous scan at 2.40°/min (0.5 sec/0.02° step) from1.5 ° 2θ to 40° 2θ was used. A sintered alumina standard was used tocheck the peak position. In general, positions of XRPD peaks areexpected to individually vary on a measurement-by-measurement basis byabout ±0.2° 2θ. In general, as understood in the art, two XRPD patternsmatch one another if the characteristic peaks of the first pattern arelocated at approximately the same positions as the characteristic peaksof the second pattern. As understood in the art, determining whether twoXRPD patterns match or whether individual peaks in two XRPD patternsmatch may require consideration of individual variables and parameterssuch as, but not limited to, preferred orientation, phase impurities,degree of crystallinity, particle size, variation in diffractometerinstrument setup, variation in XRPD data collection parameters, and/orvariation in XRPD data processing, among others. The determination ofwhether two patterns match may be performed by eye and/or by computeranalysis. Examples of XRPD patterns collected and analyzed using thesemethods and parameters are provided herein, e.g., as FIG. 1, FIG. 5,FIG. 9, FIG. 13, FIG. 17, FIG. 21 and FIG. 25.

Differential Scanning calorimetry (DSC) analyses were performed on a TAInstruments Q1000™. About 5 mg of sample was placed into a tared DSC panand the weight of the sample was accurately recorded. Typically, thesample was heated under nitrogen at a rate of about 10° C./min fromabout 25° C. up to a final temperature of about 200° C. Typically,thermal events were reported as extrapolated onset temperatures.Examples of DSC thermograms collected and analyzed using these methodsand parameters are provided herein, e.g., as FIG. 2, FIG. 6, FIG. 10,FIG. 14, FIG. 18, FIG. 22 and FIG. 26.

Thermal Gravimetric Analyses (TGA) were performed on a TA InstrumentsQ500™. Calcium oxalate was used for calibration. About 10 mg of samplewas placed on a pan, accurately weighed and loaded into the TGA furnace.The sample was heated under nitrogen at a rate of about 10° C./min fromabout 25° C. up to a final temperature of about 200° C. Examples of TGAthermograms collected and analyzed using these methods and parametersare provided herein, e.g., as FIG. 3, FIG. 7, FIG. 11, FIG. 15, FIG. 19,FIG. 23 and FIG. 27.

Solvation solvents were identified and quantified by TG-IR experimentsusing a TA Instruments Q500™ TGA interfaced with a Thermo Nicolet AEMFourier transform IR spectrophotometer. Typically a sample size of about20-50 mg was weighed into an aluminum pan and heated to about 200° C.During the TGA run, the vapor was transferred to the cell through aheated transfer line. The temperature of both transfer line and the cellwere set at about 225° C. IR spectra were collected every 10-secondrepeat time. Volatiles were identified from a search of the Aldrichvapor phase spectral library and the library match results are presentedto show the identified vapor.

Morphology and particle size analysis of the samples were carried outusing an Olympus microscope. The instrument was calibrated with USPstandards. D₉₀ values were determined using the software ImagePlus—Material Plus. The D₉₀ value represents the 90th percentile of theparticle size distribution as measured by length; i.e., 90% of theparticles have a length of this value or less.

5.12.3. Solid Form Screening Study Results

Solid forms comprising Compound A which were prepared during the solidform screening studies included Forms A, B, C, D, E, F, G and anamorphous form. Representative XRPD patterns, DSC plots, TGA plots andDVS plots for each of Forms A, B, C, D, E, F and G are provided hereinas FIG. 1-FIG. 28.

Solubility Studies.

The approximate solubility of Form B of Compound A in various solventsat about 25° C. was determined. Results are shown in Table 6. Form B wasfound to be most soluble in acetone, acetonitrile, methylene chloride,methyl ethyl ketone and tetrahydrofuran (greater than about 50 mg/mL)followed by ethyl acetate (about 30.15 mg/mL). Form B was also found tohave low solubility in several solvents including n-butanol, heptane,2-propanol, toluene and water (less than about 1 mg/mL).

Solution Evaporation Studies.

Results from solution evaporation studies performed at about 25° C. andabout 50° C. are summarized in Table 7.

Equilibration Studies.

Results from equilibration studies performed at about 25° C. and about50° C. are summarized in Table 8.

Cooling Crystallization Studies.

Results from cooling crystallization studies are summarized in Table 9.Cooling crystallization studies yielded crystalline material fromnumerous solvents, including acetone, acetonitrile, n-butyl acetate,ethyl acetate, methanol, methylene chloride, methyl ethyl ketone (MEK)and tetrahydrofuran (THF). The crystalline materials obtained weretypically characterized by XRPD, DSC and TGA.

Solvent/Anti-Solvent Precipitation Studies.

Results from solvent/anti-solvent precipitation studies are summarizedin Table 10. When heptane, water and toluene were added to Form B in THFsolution at about 40° C., precipitates formed immediately. When heptane,methyl t-butyl ether (MTBE), toluene and water were added to Form B inacetonitrile solution separately at about 25° C., either a clearsolution or a mixture formed. Crystalline material fromMTBE/acetonitrile, water/acetonitrile and toluene/acetonitrile wasobtained after stirring overnight. However, no crystallization occurredfor heptane/acetonitrile mixture. When water was added to Form B inmethanol solution at about 50° C., precipitates formed immediately andwhen heptane and toluene were added to Form B in methanol solutionseparately at about 50° C., either a clear solution or a mixture formed.Crystalline material from toluene/methanol and heptane/methanol wasobtained after stirring overnight. When toluene was added to Form B inmethylene chloride solution at about 25° C., precipitates formedimmediately and when MTBE was added to Form B in methylene chloridesolution at about 25° C., a clear solution was obtained. Crystallinematerial from MTBE/methylene chloride was obtained after stirredovernight. However, no crystallization occurred when heptane was addedto Form B in methylene chloride solution. When heptane was added to FormB in MEK solution at about 50° C., precipitates formed immediately andwhen MTBE and toluene were added to Form B in MEK solution separately atabout 50° C., clear solutions were obtained. Crystalline material fromMTBE/MEK and toluene/MEK was obtained after stirring overnight. Whenheptane was added to Form B in n-butyl acetate solution at about 50° C.,precipitates formed immediately and when MTBE and toluene were added toForm B in MEK solution separately at about 50° C., clear solutions wereobtained. Crystalline material from MTBE/n-butyl acetate andtoluene/n-butyl acetate was obtained after stirring overnight. Whenwater and toluene were added to Form B in acetone solution separately atabout 40° C., precipitates formed immediately and when ethanol and2-propanol were added to Form B in acetone solution separately at about40° C., clear solutions were obtained. Crystalline material fromethanol/acetone and 2-propanol/acetone were obtained after stirringovernight. Crystalline materials obtained were identified by XRPD, DSC,TGA.

Stability Studies.

Stability study results are summarized in Table 11. The stabilities ofForms A, B, C and D were studied by exposing the solid samples to thestress condition of 40° C./75% RH for four weeks. Moreover, thestabilities of Forms A, B, C and D in different solvents were studied byequilibration in different solvents at 40° C. for four weeks. Theslurries then were filtered and dried in the air. Solid samples obtainedfrom the stability experiments were analyzed by XRPD and DSC.

Interconversion Studies.

Results from interconversion studies are summarized in Table 12.

Compression Studies.

Compression tests were performed on Forms A, B, C, D, E, F and G ofCompound A. Each form studied was found to be substantially physicallystable as observed by XRPD analysis.

Hygroscopicity Studies.

Hygroscopicity (moisture sorption/desorption) studies were performed onForms A, B, C, D, E, F and G. Each of the solid samples were analyzed byXRPD after undergoing a full adsorption/desorption cycle in the DVSsystem. XRPD results indicated that none of the forms analyzed underwentsubstantial solid-state transformation as a result of DVS analysis.

TABLE 6 Solubility Study on Form B Approximate Solubility Solvent System(mg/ml) Acetone >50 Acetonitrile >50 n-Butanol >0.72 n-Butyl acetate9.75 Absolute ethanol 1.38 Ethyl acetate 30.15 Heptane 0.41 Methylenechloride >50 Methyl ethyl ketone >50 Methanol 4.05 Methyl t-butyl ether1.17 2-Propanol 0.81 Tetrahydrofuran >50 Toluene 0.90 Water 0.69Ethanol:Water (1:1) 2.86

TABLE 7 Solution Evaporation Studies Starting Evaporation XRPD DSCthermal Form Solvent System Temp. (° C.) Analysis events B Acetone 25Form B B Acetonitrile 25 Form B + 77.28° C.; Form E 151.84° C. B n-Butylacetate 25 Form B B Ethyl acetate 25 Form B B Methylene chloride 25 FormD 93.11° C. B Methyl ethyl ketone 25 Form B B Tetrahydrofuran 25 Form BB Ethanol:Water (1:1) 25 Form B A Acetonitrile 25 Form E 95.42° C. (TGAwt. loss = 3.56%) A Methylene chloride 25 Form D 97.23° C. A Acetone 50Form B A Acetonitrile 50 Form B A n-Butyl acetate 50 Form B A Ethylacetate 50 Form B A Methyl ethyl ketone 50 Form B A Tetrahydrofuran 50Form B A Ethanol:Water (1:1) 50 Form B

TABLE 8 Equilibration Studies Equilib. Starting Temp. XRPD DSC ThermalForm Solvent System ° C. Analysis Events B n-Butanol 25 Form B B n-Butylacetate 25 Form B B Ethanol 25 Form B B Ethyl acetate 25 Form B BHeptane 25 Form B B Methanol 25 Form B B Methyl t-butyl ether 25 Form BB 2-Propanol 25 Form B B Toluene 25 Form C 159.31° C. B Toluene (evap.25 Form C Broad multiplet at 60° C.) B Toluene:Acetone (9:1) 25 Form CBroad multiplet (evap. at 100° C.) (TGA wt. loss = 5.90%) B Water 25Form B B Water (50 days) 25 Form B A Ethanol 25 Form F 145.06° C.(multiplet) A Heptane 25 Form A A Ethyl acetate 25 Form G 108.96° C. AWater 25 Form A A Toluene 25 Form C 170.18° C. (TGA wt. loss = 5.86%) AToluene (evap. 25 Form C 167.84° C. at 60° C.) A Toluene:Acetone (9:1)25 Form C Broad multiplet (evap. at 100° C.) A Acetone:Ethanol (1:1) 25Form B 154.00° C. (main) A Ethanol:Water (1:1) 25 Form F 145.22° C. An-Butanol 50 Form B A n-Butyl acetate 50 Form B A Ethanol 50 Form B AHeptane 50 Form B A Methanol 50 Form B A Methyl t-butyl ether 50 Form BA 2-Propanol 50 Form B A Toluene 50 Form C 165.30° C. (multiplet) AWater 50 Form B A Ethanol:Water (1:1) 50 Form B

TABLE 9 Cooling Crystallization Studies Starting Analysis DSC ThermalForm Solvent System by XRPD Events B Acetone Form E B Acetonitrile FormE  95.42° C. B n-Butyl acetate Form B B Ethyl acetate Form B B MethyleneChloride Form D 100.90° C. B Methanol Form B B Methyl ethyl ketone FormB B THF Form H

TABLE 10 Solvent/Anti-Solvent Precipitation Studies Starting Anti- Ratio(Solvent: Analysis by DSC Thermal Form Solvent* Solvent* Antisolvent) &Temp. XRPD Events B Acetone Ethanol 1:8 at 40° C. Form B B Acetone2-Propanol 1:10 at 40° C. Form B B Acetone Water 1:4 at 40° C. Form B BAcetone Toluene 1:10 at 40° C. Form C 167.57° C. (broad) B AcetonitrileHeptane 1:8 at 25° C. Form B B Acetonitrile MtBE 1:8 at 25° C. Form B BAcetonitrile Water 1:6 at 25° C. Form B B Acetonitrile Toluene 1:8 at50° C. Form C 167.97° C. B Methyl ethyl Heptane 1:3 at 50° C. Form Bketone B MEK MtBE 1:4 at 50° C. Form B B MEK Toluene 1:3 at 50° C. FormC 168.22° C. B n-Butyl acetate Heptane 1:4 at 50° C. Form B B n-Butylacetate MtBE 1:4 at 50° C. Form B B n-Butyl acetate Toluene 1:4 at 50°C. Form B B DCM Heptane 1:8 at 25° C. Form E + B 89.65° C.; 149.81° C. BDCM MtBE 1:15 at 25° C. Form B B DCM Toluene 1:15 at 25° C. Form B167.99° C. (multiplet) B Methanol Heptane 1:3 at 50° C. Form B BMethanol Water 1:3 at 50° C. Form B B Methanol Toluene 1:3 at 50° C.Form C 168.37° C. (multiplet) B Tetrahydrofuran Heptane 1:6 at 40° C.Form B B Tetrahydrofuran Water 1:6 at 40° C. Form B B TetrahydrofuranToluene 1:6 at 40° C. Form C 168.64° C. (multiplet) *Abbreviations: MEK= methyl ethyl ketone; DCM = dichloromethane (i.e., methylene chloride);MtBE = methyl t-butyl ether

TABLE 11 Stability Studies Test Conditions Starting (“EQ” = equilibrate;Analysis Form “RH” = relative humidity) Appearance by XRPD Form A 40°C./75% RH; 4 weeks White solid Form A Form B 40° C./75% RH; 4 weeksWhite solid Form B Form C 40° C./75% RH; 4 weeks Yellow solid Form CForm D 40° C./75% RH; 4 weeks White solid Form D Form A EQ in ethanol at40° C. for 4 weeks Form F Form A EQ in heptane at 40° C. for 4 weeksForm A Form A EQ in water at 40° C. for 4 weeks Form A Form A EQ intoluene at 40° C. for 4 weeks Form C Form B EQ in ethanol at 40° C. for4 weeks Form B Form B EQ in heptane at 40° C. for 4 weeks Form B Form BEQ in water at 40° C. for 4 weeks Form B Form B EQ in toluene at 40° C.for 4 weeks Form B Form C EQ in ethanol at 40° C. for 4 weeks Form CForm C EQ in heptane at 40° C. for 4 weeks Form C Form C EQ in water at40° C. for 4 weeks Form C Form C EQ in toluene at 40° C. for 4 weeksForm C Form D EQ in ethanol at 40° C. for 4 weeks Form B Form D EQ inheptane at 40° C. for 4 weeks Form B Form D EQ in water at 40° C. for 4weeks Form B Form D EQ in toluene at 40° C. for 4 weeks Form C

TABLE 12 Interconversion Studies Test Conditions Analysis Starting Form(“EQ” = equilibrate) by XRPD Mixture of Forms EQ in acetone:ethanol(1:1) at 25° C. Form B + A, B, C, D, C + F E, F and G Form A EQ inacetone:ethanol (1:1) at 25° C. Form B Form C EQ in acetone:ethanol(1:1) at 25° C. Form C Form D EQ in acetone:ethanol (1:1) at 25° C. FormB Form E EQ in acetone:ethanol (1:1) at 25° C. Form B Form F EQ inacetone:ethanol (1:1) at 25° C. Form F Form G EQ in acetone:ethanol(1:1) at 25° C. Form B

5.13. Example 13 200 mg Dosage Capsule

Table 13 illustrates a batch formulation and single dosage formulationfor a single dose unit containing 200 mg of a solid form comprisingCompound A, i.e., about 40 percent by weight, in a size #0 capsule.

TABLE 13 Formulation for 200 mg capsule Percent Quantity QuantityMaterial By Weight (mg/tablet) (kg/batch) Compound A 40.0% 200 mg 16.80kg Pregelatinized Corn 9.5% 297.5 mg 24.99 kg Starch, NF5 MagnesiumStearate 0.5% 2.5 mg 0.21 kg Total 100.0% 500 mg 42.00 kg

The pregelatinized corn starch (SPRESS™ B-820) and Compound A componentsare passed through a 710 μm screen and then are loaded into a DiffusionMixer with a baffle insert and blended for 15 minutes. The magnesiumstearate is passed through a 210 μm screen and is added to the DiffusionMixer. The blend is then encapsulated in a size #0 capsule, 500 mg percapsule (8400 capsule batch size) using a Dosator type capsule fillingmachine

5.14. Example 14 100 mg Oral Dosage Form

Table 14 illustrates a batch formulation and a single dose unitformulation containing 100 mg of a solid form comprising Compound A.

TABLE 14 Formulation for 100 mg tablet Percent Quantity QuantityMaterial by Weight (mg/tablet) (kg/batch) Compound A  40% 100.00 20.00Microcrystalline 53.5%  133.75 26.75 Cellulose, NF Pluronic F-68 4.0%10.00 2.00 Surfactant Croscarmellose 2.0% 5.00 1.00 Sodium Type A, NFMagnesium Stearate, 0.5% 1.25 0.25 NF Total 100.0%  250.00 mg 50.00 kg

The microcrystalline cellulose, croscarmellose sodium, and Compound Acomponents are passed through a #30 mesh screen (about 430μ to about655μ). The Pluronic F-68® (manufactured by JRH Biosciences, Inc. ofLenexa, Kans.) surfactant is passed through a #20 mesh screen (about457μ to about 1041μ). The Pluronic F-68® surfactant and 0.5 kgs ofcroscarmellose sodium are loaded into a 16 qt. twin shell tumble blenderand are mixed for about 5 minutes. The mix is then transferred to a 3cubic foot twin shell tumble blender where the microcrystallinecellulose is added and blended for about 5 minutes. The solid formcomprising Compound A is added and blended for an additional 25 minutes.This pre-blend is passed through a roller compactor with a hammer millattached at the discharge of the roller compactor and moved back to thetumble blender. The remaining croscarmellose sodium and magnesiumstearate is added to the tumble blender and blended for about 3 minutes.The final mixture is compressed on a rotary tablet press with 250 mg pertablet (200,000 tablet batch size).

While the invention has been described with respect to the particularembodiments, it will be apparent to those skilled in the art thatvarious changes and modifications may be made without departing from thespirit and scope of the invention as defined in the claims. Suchmodifications are also intended to fall within the scope of the appendedclaims.

1-39. (canceled)
 40. A method of treating or preventing a disease ordisorder ameliorated by the inhibition of TNF-α production, wherein themethod comprises administering a therapeutically or prophylacticallyeffective amount of a Form B crystal form of the compound of Formula(I):

which is enantiomerically pure, and which has an X-ray powderdiffraction pattern comprising peaks at about 10.1, 13.5, 20.7, and 26.9degrees 2θ.
 41. The method of claim 40, wherein the crystal form has anX-ray powder diffraction pattern further comprising peaks at about 12.4,15.7, 18.1, and 24.7 degrees 2θ.
 42. The method of claim 41, wherein thecrystal form has an X-ray powder diffraction pattern further comprisingpeaks at about 16.3, 22.5, 26.2, and 29.1 degrees 2θ.
 43. The method ofclaim 40, wherein the crystal form has an X-ray powder diffractionpattern matching the pattern depicted in FIG.
 5. 44. The method of claim40, wherein the crystal form has a differential scanning calorimetryplot comprising an endothermic event with an onset temperature of about154° C.
 45. The method of claim 40, wherein the crystal form has adifferential scanning calorimetry plot matching the plot depicted inFIG.
 6. 46. The method of claim 40, wherein the crystal form has athermal gravimetric analysis plot comprising a mass loss of less thanabout 1% when heated from about 25° C. to about 140° C.
 47. The methodof claim 46, wherein the mass loss is about 0.25%.
 48. The method ofclaim 40, wherein the crystal form has a thermal gravimetric analysisplot matching the plot depicted in FIG.
 7. 49. The method of claim 40,wherein the crystal form exhibits a mass increase of less than about 1%when subjected to an increase in relative humidity from about 0% toabout 95% relative humidity.
 50. The method of claim 49, wherein themass increase is about 0.6%.
 51. The method of claim 40, wherein thecrystal form has a moisture sorption isotherm plot matching the plotdepicted in FIG.
 8. 52. The method of claim 40, wherein the crystal formis stable upon exposure to about 40° C. and about 75% relative humidityfor about 4 weeks.
 53. The method of claim 40, wherein the disease ordisorder ameliorated by the inhibition of TNF-α production is psoriasis.54. The method of claim 40, wherein the disease or disorder amelioratedby the inhibition of TNF-α production is psoriatic arthritis.
 55. Themethod of claim 40, wherein the disease or disorder ameliorated by theinhibition of TNF-α production is rheumatoid arthritis.
 56. The methodof claim 40, wherein the disease or disorder ameliorated by theinhibition of TNF-α production is Behcet's Disease.
 57. The method ofclaim 40, wherein the disease or disorder ameliorated by the inhibitionof TNF-α production is rheumatoid spondylitis.
 58. The method of claim40, wherein the disease or disorder ameliorated by the inhibition ofTNF-α production is an arthritic condition.